Tricyclic p13k inhibitor compounds and methods of use

ABSTRACT

Tricyclic PI3k inhibitor compounds of Formula I with anti-cancer activity, anti-inflammatory activity, or immunoregulatory properties, and more specifically with PI3 kinase modulating or inhibitory activity are described. Methods are described for using the tricyclic PI3K inhibitor compounds of Formula I for in vitro, in situ, and in vivo diagnosis or treatment of mammalian cells, organisms, or associated pathological conditions. 
     
       
         
         
             
             
         
       
     
     Formula I compounds include stereoisomers, geometric isomers, tautomers, and pharmaceutically acceptable salts thereof. The dashed lines indicate an optional double bond, and at least one dashed line is a double bond. The substituents are as described.

CROSS REFERENCE TO RELATED APPLICATIONS

This non-provisional application filed under 37 CFR § 1.53(b), claimsthe benefit under 35 USC § 119(c) of U.S. Provisional Application Ser.No. 61/423,694 filed on 16 Dec. 2010, which is incorporated by referencein entirety

FIELD OF THE INVENTION

The invention relates generally to compounds with anti-cancer activityor anti-inflammatory activity, and more specifically to compounds whichinhibit PI3 kinase activity. The invention also relates to methods ofusing the compounds for in vitro, in situ, and in vivo diagnosis ortreatment of mammalian cells, or associated pathological conditions.

BACKGROUND OF THE INVENTION

Phosphatidylinositol is one of a number of phospholipids found in cellmembranes which play an important role in intracellular signaltransduction. Cell signaling via 3′-phosphorylated phosphoinositides hasbeen implicated in a variety of cellular processes, e.g., malignanttransformation, growth factor signaling, inflammation, and immunity(Rameh et al (1999) J. Biol Chem, 274:8347-8350). The enzyme responsiblefor generating these phosphorylated signaling products,phosphatidylinositol 3-kinase (also referred to as PI 3-kinase or PI3K),was originally identified as an activity associated with viraloncoproteins and growth factor receptor tyrosine kinases thatphosphorylate phosphatidylinositol (PI) and its phosphorylatedderivatives at the 3′-hydroxyl of the inositol ring (Panayotou et at(1992) Trends Cell Biol 2:358-60).

Phosphoinositide 3-kinases (PI3K) are lipid kinases that phosphorylatelipids at the 3-hydroxyl residue of an inositol ring (Whitman et al(1988) Nature, 332:664). The 3-phosphorylated phospholipids (PIP3s)generated by PI3-kinases act as second messengers recruiting kinaseswith lipid binding domains (including plekstrin homology (PH) regions),such as Akt and phosphoinositide-dependent kinase-1 (PDK1). Binding ofAkt to membrane PIP3s causes the translocation of Akt to the plasmamembrane, bringing Akt into contact with PDK1, which is responsible foractivating Akt. The tumor-suppressor phosphatase, PTEN, dephosphorylatesPIP3 and therefore acts as a negative regulator of Akt activation. ThePI3-kinases Akt and PDK1 are important in the regulation of manycellular processes including cell cycle regulation, proliferation,survival, apoptosis and motility and are significant components of themolecular mechanisms of diseases such as cancer, diabetes and immuneinflammation (Vivanco et al (2002) Nature Rev. Cancer 2:489; Phillips etal (1998) Cancer 83:41).

The main PI3-kinase isoform in cancer is the Class I PI3-kinase, p110 α(alpha) (U.S. Pat. Nos. 5,824,492; 5,846,824; 6,274,327). Other isoformsare implicated in cardiovascular and immune-inflammatory disease(Workman P (2004) Biochem Soc Trans 32:393-396; Patel et al (2004)Proceedings of the American Association of Cancer Research (AbstractLB-247) 95th Annual Meeting, March 27-31, Orlando, Fla., USA; Ahmadi Kand Waterfield M D (2004) Encyclopedia of Biological Chemistry (LennarzW J, Lane M D eds) Elsevier/Academic Press). The PI3 kinase/Akt/PTENpathway is an attractive target for cancer drug development since suchmodulating or inhibitory agents would be expected to inhibitproliferation, reverse the repression of apoptosis and surmountresistance to cytotoxic agents in cancer cells (Folkes et al (2008) J.Med. Chem. 51:5522-5532; Yaguchi et al (2006) Jour. of the Nat. CancerInst. 98(8):545-556).

Malignant gliomas are the most common primary brain tumors in adults. Inglioblastoma (GBM), the most aggressive glioma subtype, tumor formationand growth appear to be driven by amplification or overexpression ofgene products involved in growth factor-initiated signal transductionacting in cooperation with genetic alterations disrupting cell-cyclecontrol (Holland EC (2001) Nat Rev Genet 2:120-129). Of the genomicalterations described in GBM, PTEN mutation and/or deletion is the mostcommon, with an estimated frequency of 70-90% (Nutt C, Louis D N (2005)Cancer of the Nervous System (McGraw-Hill, New York), 2nd Ed, pp837-847.). These findings, along with the prognostic value of PTENstatus in GBM cases (Phillips H S, et al. (2006) Cancer Cell 9:157-163),suggest the importance of the phosphoinositide 3-kinase (PI3K)/Aktpathway in promoting highly aggressive glial malignancies, as well asthe opportunities for treatment with PI3K inhibitors possessingblood-brain barrier penetrant properties.

Malignant gliomas are treated with a combination of surgery, radiation,and temozolomide (TEMODAR™), but these therapies ultimately fail at ahigh frequency due to tumor recurrence. Additional therapies are neededfor delivery of effective concentrations of effective drugs to the brainto treat hyperproliferative disorders, such as glioblastoma andmetastatic brain cancer.

SUMMARY OF THE INVENTION

The invention relates generally to tricyclic PI3K inhibitor compounds ofFormula I with anti-cancer activity, anti-inflammatory activity, orimmunoregulatory properties, and more specifically with PI3 kinasemodulating or inhibitory activity. Certain hyperproliferative disordersare characterized by the modulation of PI3 kinase function, for exampleby mutations or overexpression of the proteins. Accordingly, thecompounds of the invention may be useful in the treatment ofhyperproliferative disorders such as cancer. The compounds may inhibittumor growth in mammals and may be useful for treating human cancerpatients.

The invention also relates to methods of using the tricyclic PI3Kinhibitor compounds of Formula I for in vitro, in situ, and in vivodiagnosis or treatment of mammalian cells, organisms, or associatedpathological conditions.

Formula I compounds include:

and stereoisomers, geometric isomers, tautomers, and pharmaceuticallyacceptable salts thereof. The dashed lines indicate an optional doublebond, and at least one dashed line is a double bond. The substituentsare as described herein.

Another aspect of the invention provides a pharmaceutical compositioncomprising a tricyclic PI3K inhibitor compound of Formula I and apharmaceutically acceptable carrier. The pharmaceutical composition mayfurther comprise one or more additional therapeutic agent.

Another aspect of the invention provides methods of inhibiting PI3kinase activity, comprising contacting a PI3 kinase with an effectiveinhibitory amount of a compound of Formula I.

Another aspect of the invention provides methods of preventing ortreating a hyperproliferative disease or disorder modulated by PI13kinases, comprising administering to a mammal in need of such treatmentan effective amount of a compound of Formula I. Examples of suchhyperproliferative disease or disorder include, but are not limited to,cancer.

Another aspect of the invention provides methods of preventing ortreating a hyperproliferative disorder, comprising administering to amammal in need of such treatment an effective amount of a compound ofFormula I, alone or in combination with one or more additional compoundshaving anti-hyperproliferative properties.

In a further aspect the present invention provides a method of using acompound of this invention to treat a hyperproliferative disease orcondition modulated by PI3 kinase in a mammal.

An additional aspect of the invention is the use of a compound of thisinvention for treating cancer modulated by PI3 kinase in a mammal.

Another aspect of the invention includes kits comprising a compound ofFormula I, a container, and optionally a package insert or labelindicating a treatment.

Another aspect of the invention includes methods of preparing, methodsof separating, and methods of purifying compounds of Formula I.

Another aspect of the invention includes novel intermediates useful forpreparing Formula I compounds.

DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS

Reference will now be made in detail to certain embodiments of theinvention, examples of which are illustrated in the accompanyingstructures and formulas. While the invention will be described inconjunction with the enumerated embodiments, it will be understood thatthey are not intended to limit the invention to those embodiments. Onthe contrary, the invention is intended to cover all alternatives,modifications, and equivalents which may be included within the scope ofthe present invention as defined by the claims.

One skilled in the art will recognize many methods and materials similaror equivalent to those described herein, which could be used in thepractice of the present invention. The present invention is in no waylimited to the methods and materials described. In the event that one ormore of the incorporated literature, patents, and similar materialsdiffers from or contradicts this application, including but not limitedto defined terms, term usage, described techniques, or the like, thisapplication controls.

Definitions

The term “alkyl” as used herein refers to a saturated linear orbranched-chain monovalent hydrocarbon radical of one to twelve carbonatoms (C₁-C₁₂), wherein the alkyl radical may be optionally substitutedindependently with one or more substituents described below. In anotherembodiment, an alkyl radical is one to eight carbon atoms (C₁-C₅), orone to six carbon atoms (C₁-C₆). Examples of alkyl groups include, butare not limited to, methyl (Me, —CH₃), ethyl (Et, —CH₂CH₃), 1-propyl(n-Pr, n-propyl, —CH₂CH₂CH₃), 2-propyl (i-Pr, i-propyl, —CH(CH₃)₂),1-butyl (n-Bu, n-butyl, —CH₂CH₂CH₂CH₃), 2-methyl-1-propyl (i-Bu,i-butyl, —CH₂CH(CH₃)₂), 2-butyl (s-Bu, s-butyl, —CH(CH₃)CH₂CH₃),2-methyl-2-propyl (t-Bu, t-butyl, —C(CH₃)₃), 1-pentyl (n-pentyl,—CH₂CH₂CH₂CH₂CH₃), 2-pentyl (—CH(CH₃)CH₂CH₂CH₃), 3-pentyl(—CH(CH₂CH₃)₂), 2-methyl-2-butyl (—C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(—CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl (—CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(—CH₂CH(CH₃)CH₂CH₃), 1-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl(—CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl (—CH(CH₂CH₃)(CH₂CH₂CH₃)),2-methyl-2-pentyl (—C(CH₃)₂CH₂CH₂CH₃), 3-methyl-2-pentyl(—CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl (—CH(CH₃)CH₂CH(CH₃)₂),3-methyl-3-pentyl (—C(CH₃)(CH₂CH₃)₂), 2-methyl-3-pentyl(—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl (—C(CH₃)₂CH(CH₃)₂),3,3-dimethyl-2-butyl (—CH(CH₃)C(CH₃)₃, 1-heptyl, 1-octyl, and the like.

The term “alkylene” as used herein refers to a saturated linear orbranched-chain divalent hydrocarbon radical of one to twelve carbonatoms (C₁-C₁₂), wherein the alkylene radical may be optionallysubstituted independently with one or more substituents described below.In another embodiment, an alkylene radical is one to eight carbon atoms(C₁-C₈), or one to six carbon atoms (C₁-C₆). Examples of alkylene groupsinclude, but are not limited to, methylene (—CH₂—), ethylene (—CH₂CH₂—),propylene (—CH₂CH₂CH₂—), and the like.

The term “alkenyl” refers to linear or branched-chain monovalenthydrocarbon radical of two to eight carbon atoms (C₂-C₈) with at leastone site of unsaturation, i.e., a carbon-carbon, sp² double bond,wherein the alkenyl radical may be optionally substituted independentlywith one or more substituents described herein, and includes radicalshaving “cis” and “trans” orientations, or alternatively, “E” and “Z”orientations. Examples include, but are not limited to, ethylenyl orvinyl (—CH═CH₂), allyl (—CH₂CH═CH₂), and the like.

The term “alkenylene” refers to linear or branched-chain divalenthydrocarbon radical of two to eight carbon atoms (C₂-C₈) with at leastone site of unsaturation, i.e., a carbon-carbon, sp² double bond,wherein the alkenyl radical may be optionally substituted, and includesradicals having “cis” and “trans” orientations, or alternatively, “E”and “Z” orientations. Examples include, but are not limited to,ethylenylene or vinylene (—CH═CH—), allyl (—CH₂CH═CH—), and the like.

The term “alkynyl” refers to a linear or branched monovalent hydrocarbonradical of two to eight carbon atoms (C₂-C₈) with at least one site ofunsaturation, i.e., a carbon-carbon, sp triple bond, wherein the alkynylradical may be optionally substituted independently with one or moresubstituents described herein. Examples include, but are not limited to,ethynyl (—C≡CH), propynyl (propargyl, —CH₂C≡CH), and the like.

The term “alkynylene” refers to a linear or branched divalenthydrocarbon radical of two to eight carbon atoms (C₂-C₈) with at leastone site of unsaturation, i.e., a carbon-carbon, sp triple bond, whereinthe alkynyl radical may be optionally substituted independently with oneor more substituents described herein. Examples include, but are notlimited to, ethynylene (—C≡C—), propynylene (propargylene, —CH₂C≡C—),and the like.

The terms “carbocycle”, “carbocyclyl”, “carbocyclic ring” and“cycloalkyl” refer to a monovalent non-aromatic, saturated or partiallyunsaturated ring having 3 to 12 carbon atoms (C₃-C₁₂) as a monocyclicring or 7 to 12 carbon atoms as a bicyclic ring. Bicyclic carbocycleshaving 7 to 12 atoms can be arranged, e.g., as a bicyclo [4,5], [5,5],[5,6] or [6,6] system, and bicyclic carbocycles having 9 or 10 ringatoms can be arranged as a bicyclo [5,6] or [6,6] system, or as bridgedsystems such as bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane andbicyclo[3.2.2]nonane. Examples of monocyclic carbocycles include, butare not limited to, cyclopropyl, cyclobutyl, cyclopentyl,1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl,1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl,cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl,cycloundecyl, cyclododecyl, and the like.

“Aryl” means a monovalent aromatic hydrocarbon radical of 6-20 carbonatoms (C₆-C₂₀) derived by the removal of one hydrogen atom from a singlecarbon atom of a parent aromatic ring system. Some aryl groups arerepresented in the exemplary structures as “Ar”. Aryl includes bicyclicradicals comprising an aromatic ring fused to a saturated, partiallyunsaturated ring, or aromatic carbocyclic ring. Typical aryl groupsinclude, but are not limited to, radicals derived from benzene (phenyl),substituted benzenes, naphthalene, anthracene, biphenyl, indenyl,indanyl, 1,2-dihydronaphthalene, 1,2,3,4-tetrahydronaphthyl, and thelike. Aryl groups are optionally substituted independently with one ormore substituents described herein.

“Arylene” means a divalent aromatic hydrocarbon radical of 6-20 carbonatoms (C₆-C₂₀) derived by the removal of two hydrogen atom from a twocarbon atoms of a parent aromatic ring system. Some arylene groups arerepresented in the exemplary structures as “Ar”. Arylene includesbicyclic radicals comprising an aromatic ring fused to a saturated,partially unsaturated ring, or aromatic carbocyclic ring. Typicalarylene groups include, but are not limited to, radicals derived frombenzene (phenylene), substituted benzenes, naphthalene, anthracene,biphenylene, indenylene, indenylene, 1,2-dihydronaphthalene,1,2,3,4-tetrahydronaphthyl, and the like. Arylene groups are optionallysubstituted independently with one or more substituents describedherein.

The terms “heterocycle,” “heterocyclyl” and “heterocyclic ring” are usedinterchangeably herein and refer to a saturated or a partiallyunsaturated (i.e., having one or more double and/or triple bonds withinthe ring) carbocyclic radical of 3 to about 20 ring atoms in which atleast one ring atom is a heteroatom selected from nitrogen, oxygen,phosphorus and sulfur, the remaining ring atoms being C, where one ormore ring atoms is optionally substituted independently with one or moresubstituents described below. A heterocycle may be a monocycle having 3to 7 ring members (2 to 6 carbon atoms and 1 to 4 heteroatoms selectedfrom N, O, P, and S) or a bicycle having 7 to 10 ring members (4 to 9carbon atoms and 1 to 6 heteroatoms selected from N, O, P, and S), e.g.:a bicyclo [4,5], [5,5], [5,6], or [6,6] system. Heterocycles aredescribed in Paquette, Leo A.; “Principles of Modern HeterocyclicChemistry” (W. A. Benjamin, New York, 1968), particularly Chapters 1, 3,4, 6, 7, and 9; “The Chemistry of Heterocyclic Compounds, A series ofMonographs” (John Wiley & Sons, New York, 1950 to present), inparticular Volumes 13, 14, 16, 19, and 28; and J. Am. Chem. Soc. (1960)82:5566. “Heterocyclyl” also includes radicals where heterocycleradicals are fused with a saturated, partially unsaturated ring, oraromatic carbocyclic or heterocyclic ring. Examples of heterocyclicrings include, but are not limited to, morpholin-4-yl, piperidin-1-yl,piperazinyl, piperazin-4-yl-2-one, piperazin-4-yl-3-one,pyrrolidin-1-yl, thiomorpholin-4-yl, S-dioxothiomorpholin-4-yl,azocan-1-yl, azetidin-1-yl, octahydropyrido[1,2-a]pyrazin-2-yl,[1,4]diazepan-1-yl, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl,tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino,thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl,thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl,thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl,4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl,dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl,pyrazolidinylimidazolinyl, imidazolidinyl, 3-azabicyco[3.1.0]hexanyl,3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 3H-indolylquinolizinyl and N-pyridyl ureas. Spiro moieties are also includedwithin the scope of this definition. Examples of a heterocyclic groupwherein 2 ring atoms are substituted with oxo (═O) moieties arepyrimidinonyl and 1,1-dioxo-thiomorpholinyl. The heterocycle groupsherein are optionally substituted independently with one or moresubstituents described herein.

The term “heteroaryl” refers to a monovalent aromatic radical of 5-, 6-,or 7-membered rings, and includes fused ring systems (at least one ofwhich is aromatic) of 5-20 atoms, containing one or more heteroatomsindependently selected from nitrogen, oxygen, and sulfur. Examples ofheteroaryl groups are pyridinyl (including, e.g., 2-hydroxypyridinyl),imidazolyl, imidazopyridinyl, pyrimidinyl (including, e.g.,4-hydroxypyrimidinyl), pyrazolyl, triazolyl, pyrazinyl, tetrazolyl,furyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl,isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl,tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl,cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl,triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, triazolyl,thiadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl,naphthyridinyl, and furopyridinyl. Heteroaryl groups are optionallysubstituted independently with one or more substituents describedherein.

The heterocycle or heteroaryl groups may be carbon (carbon-linked), ornitrogen (nitrogen-linked) bonded where such is possible. By way ofexample and not limitation, carbon bonded heterocycles or heteroarylsare bonded at position 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5,or 6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position2, 3, 5, or 6 of a pyrazine, position 2, 3, 4, or 5 of a furan,tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole,position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4,or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of anaziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6,7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of anisoquinoline.

By way of example and not limitation, nitrogen bonded heterocycles orheteroaryls are bonded at position 1 of an aziridine, azetidine,pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole,imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline,2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline,1H-indazole, position 2 of a isoindole, or isoindoline, position 4 of amorpholine, and position 9 of a carbazole, or β-carboline.

The terms “treat” and “treatment” refer to both therapeutic treatmentand prophylactic or preventative measures, wherein the object is toprevent or slow down (lessen) an undesired physiological change ordisorder, such as the development or spread of cancer. For purposes ofthis invention, beneficial or desired clinical results include, but arenot limited to, alleviation of symptoms, diminishment of extent ofdisease, stabilized (i.e., not worsening) state of disease, delay orslowing of disease progression, amelioration or palliation of thedisease state, and remission (whether partial or total), whetherdetectable or undetectable. “Treatment” can also mean prolongingsurvival as compared to expected survival if not receiving treatment.Those in need of treatment include those already with the condition ordisorder as well as those prone to have the condition or disorder orthose in which the condition or disorder is to be prevented.

The phrase “therapeutically effective amount” means an amount of acompound of the present invention that (i) treats or prevents theparticular disease, condition, or disorder, (ii) attenuates,ameliorates, or eliminates one or more symptoms of the particulardisease, condition, or disorder, or (iii) prevents or delays the onsetof one or more symptoms of the particular disease, condition, ordisorder described herein. In the case of cancer, the therapeuticallyeffective amount of the drug may reduce the number of cancer cells;reduce the tumor size; inhibit (i.e., slow to some extent and preferablystop) cancer cell infiltration into peripheral organs; inhibit (i.e.,slow to some extent and preferably stop) tumor metastasis; inhibit, tosome extent, tumor growth; and/or relieve to some extent one or more ofthe symptoms associated with the cancer. To the extent the drug mayprevent growth and/or kill existing cancer cells, it may be cytostaticand/or cytotoxic. For cancer therapy, efficacy can be measured, e.g., byassessing the time to disease progression (TTP) and/or determining theresponse rate (RR).

The terms “cancer” refers to or describe the physiological condition inmammals that is typically characterized by unregulated cell growth. A“tumor” comprises one or more cancerous cells. Examples of cancerinclude, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma,and leukemia or lymphoid malignancies. More particular examples of suchcancers include squamous cell cancer (e.g., epithelial squamous cellcancer), lung cancer including small-cell lung cancer, non-small celllung cancer (“NSCLC”), adenocarcinoma of the lung and squamous carcinomaof the lung, cancer of the peritoneum, hepatocellular cancer, gastric orstomach cancer including gastrointestinal cancer, pancreatic cancer,glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladdercancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectalcancer, endometrial or uterine carcinoma, salivary gland carcinoma,kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer,hepatic carcinoma, anal carcinoma, penile carcinoma, as well as head andneck cancer.

A “chemotherapeutic agent” is a chemical compound useful in thetreatment of cancer, regardless of mechanism of action. Classes ofchemotherapeutic agents include, but are not limited to: alkyatingagents, antimetabolites, spindle poison plant alkaloids,cytoxic/antitumor antibiotics, topoisomerase inhibitors, antibodies,photosensitizers, and kinase inhibitors. Chemotherapeutic agents includecompounds used in “targeted therapy” and conventional chemotherapy.Examples of chemotherapeutic agents include: erlotinib (TARCEVA®),Genentech/OSI Pharm.), docetaxel (TAXOTERE®, Sanofi-Aventis), 5-FU(fluorouracil, 5-fluorouracil, CAS No. 51-21-8), gemcitabine (GEMZAR®,Lilly), PD-0325901 (CAS No. 391210-10-9, Pfizer), cisplatin(cis-diamine,dichloroplatinum(II), CAS No. 15663-27-1), carboplatin (CASNo. 41575-94-4), paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology,Princeton, N.J.), pemetrexed (ALIMTA®, Eli Lilly), trastuzumab(HERCEPTIN®, Genentech), temozolomide(4-methyl-5-oxo-2,3,4,6,8-pentazabicyclo [4.3.0]nona-2,7,9-triene-9-carboxamide, CAS No. 85622-93-1, TEMODAR®, TEMODAL®,Schering Plough), tamoxifen((Z)-2-[4-(1,2-diphenylbut-1-enyl)phenoxy]-N,N-dimethylethanamine,NOLVADEX®, ISTUBAL®, VALODEX®), and doxorubicin (ADRIAMYCIN®, Akti-1/2,HPPD, and rapamycin.

More examples of chemotherapeutic agents include: oxaliplatin(ELOXATIN®, Sanofi), bortezomib (VELCADE®, Millennium Pharm.), sutent(SUNITINIB®, SU11248, Pfizer), letrozole (FEMARA®, Novartis), imatinibmesylate (GLEEVEC®, Novartis), XL-518 (Mek inhibitor, Exelixis, WO2007/044515), ARRY-886 (Mek inhibitor, AZD6244, Array BioPharma, AstraZeneca), SF-1126 (PI3K inhibitor, Semafore Pharmaceuticals), BEZ-235(PI3K inhibitor, Novartis), XL-147 (PI3K inhibitor, Exelixis), PTK787/ZK222584 (Novartis), fulvestrant (FASLODEX®, AstraZeneca), leucovorin(folinic acid), rapamycin (sirolimus, RAPAMUNE®, Wyeth), lapatinib(TYKERB®, GSK572016, Glaxo Smith Kline), lonafarnib (SARASAR®, SCH66336, Schering Plough), sorafenib (NEXAVAR®, BAY43-9006, Bayer Labs),gefitinib (IRESSA®, AstraZeneca), irinotecan (CAMPTOSAR®, CPT-11,Pfizer), tipifarnib (ZARNESTRA™, Johnson & Johnson), ABRAXANE™(Cremophor-free), albumin-engineered nanoparticle formulations ofpaclitaxel (American Pharmaceutical Partners, Schaumberg, II),vandetanib (rINN, ZD6474, ZACTIMA®, AstraZeneca), chloranmbucil, AG1478,AG1571 (SU 5271; Sugen), temsirolimus (TORISEL®, Wyeth), pazopanib(GlaxoSmithKline), canfosfamide (TELCYTA®, Telik), thiotepa andcyclosphosphamide (CYTOXAN®, NEOSAR®); alkyl sulfonates such asbusulfan, improsulfan and piposulfan; aziridines such as benzodopa,carboquone, meturedopa, and uredopa; ethylenimines and methylamelaminesincluding altretamine, triethylenemelamine, triethylenephosphoramide,triethylenethiophosphoramide and trimethylomelamine; acetogenins(especially bullatacin and bullatacinone); a camptothecin (including thesynthetic analog topotecan); bryostatin; callystatin; CC-1065 (includingits adozelesin, carzelesin and bizelesin synthetic analogs);cryptophycins (particularly cryptophycin 1 and cryptophycin 8);dolastatin; duocarmycin (including the synthetic analogs, KW-2189 andCB1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin;nitrogen mustards such as chlorambucil, chlornaphazine,chlorophosphamide, estramustine, ifosfamide, mechlorethamine,mechlorethamine oxide hydrochloride, melphalan, novembichin,phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureassuch as carmustine, chlorozotocin, fotemustine, lomustine, nimustine,and ranimnustine: antibiotics such as the enediyne antibiotics (e.g.,calicheamicin, calicheamicin gamma11, calicheamicin omega11 (Angew Chem.Intl. Ed. Engl. (1994) 33:183-186); dynemicin, dynemicin A;bisphosphonates, such as elodronate; an esperamicin; as well asneocarzinostatin chromophore and related chromoprotein enediyneantibiotic chromophores), aclacinomysins, actinomycin, authramycin,azaserine, bleomycins, cactinomycin, carabicin, carminomycin,carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucince, morpholino-doxorubicin,cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin anddeoxydoxorubicin), epirubicin, esorubicin, idarubicin, nemorubicin,marcellomycin, mitomycins such as mitomycin C, mycophenolic acid,nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin,quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexateand 5-fluorouracil (5-FU); folic acid analogs such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine;androgens such as calusterone, dromostanolone propionate, epitiostanol,mepitiostane, testolactone; anti-adrenals such as aminoglutethimide,mitotane, trilostane; folic acid replenisher such as frolinic acid;aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil;amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine;diaziquone; elfornithine; elliptinium acetate; an epothilone; etoglucid;gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids suchas maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol;nitracrine; pentostatin; phenamet; pirarubicin; losoxantrone;podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharidecomplex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin;sizofiran; spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-trichlorotriethylamine; trichothecenes, verracurin A, roridin Aand anguidine); urethan; vindesine; dacarbazine; mannomustine;mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”);cyclophosphamide; thiotepa; 6-thioguanine; mercaptopurine; methotrexate;platinum analogs such as cisplatin and carboplatin; vinblastine;etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine(NAVELBINE®; novantrone; teniposide; edatrexate; daunomycin;aminopterin; capecitabine (XELODAR®, Roche); ibandronate; CPT-11;topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO);retinoids such as retinoic acid; and pharmaceutically acceptable salts,acids and derivatives of any of the above.

Also included in the definition of “chemotherapeutic agent” are: (i)anti-hormonal agents that act to regulate or inhibit hormone action ontumors such as anti-estrogens and selective estrogen receptor modulators(SERMs), including, e.g., tamoxifen (including NOLVADEX®; tamoxifencitrate), raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene,keoxifene, LY117018, onapristone, and FARESTON® (toremifine citrate);(ii) aromatase inhibitors that inhibit the enzyme aromatase, whichregulates estrogen production in the adrenal glands, such as, e.g.,4(5)-imidazoles, aminoglutethimide, MEGASE® (megestrol acetate),AROMASIN® (exemestane; Pfizer), formestanie, fadrozole, RIVISOR®(vorozole), FEMARA® (lectrozole; Novartis), and ARIMIDEX® (anastrozole;AstraZeneca); (iii) anti-androgens such as flutamide, nilutamide,bicalutamide, leuprolide, and goserelin; as well as troxacitabine (a1,3-dioxolane nucleoside cytosine analog); (iv) protein kinaseinhibitors such as MEK inhibitors (WO 2007/044515); (v) lipid kinaseinhibitors; (vi) antisense oligonucleotides, particularly those whichinhibit expression of genes in signaling pathways implicated in aberrantcell proliferation, e.g., PKC-alpha, Raf and H-Ras, such as oblimersen(GENASENSE®, Genta Inc.); (vii) ribozymes such as VEGF expressioninhibitors (e.g., ANGIOZYME®) and HER2 expression inhibitors; (viii)vaccines such as gene therapy vaccines, e.g., ALLOVECTIN®, LEUVECTIN®,and VAXID®; PROLEUKIN® rIL-2; topoisomerase 1 inhibitors such asLURTOTECAN®; ABARELIX® rmRH; (ix) anti-angiogenic agents such asbevacizumab (AVASTIN®, Genentech); and pharmaceutically acceptablesalts, acids and derivatives of any of the above.

Also included in the definition of“chemotherapeutic agent” aretherapeutic antibodies such as alemtuzumab (Campath), bevacizumab(AVASTIN®, Genentech); cetuximab (ERBITUX®, Imelone); panitumumab(VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech/Biogen Idec),pertuzumab (OMNITARG™, 2C4, Genentech), trastuzumab (HERCEPTIN®,Genentech), tositumomab (Bexxar, Corixia), and the antibody drugconjugate, gemtuzumab ozogamicin (MYLOTARG®, Wyeth).

Humanized monoclonal antibodies with therapeutic potential aschemotherapeutic agents in combination with the PI3K inhibitors of theinvention include: alemtuzumab, apolizumab, aselizumab, atlizumab,bapineuzumab, bevacizumab, bivatuzumab mertansine, cantuzumabmertansine, cedelizumab, certolizumab pegol, cidfusituzumab, cidtuzumab,daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab,fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab,labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab,motovizumab, natalizumab, nimotuzumab, nolovizumab, numavizumab,ocrelizumab, omalizumab, palivizumab, pascolizumab, pecfusituzumab,pectuzumab, pertuzumab, pexelizumab, ralivizumab, ranibizumab,reslivizumab, reslizumab, resyvizumab, rovelizumab, ruplizumab,sibrotuzumab, siplizumab, sontuzumab, tacatuzumab tetraxetan,tadocizumab, talizumab, tefibazumab, tocilizumab, toralizumab,trastuzumab, tucotuzumab celmoleukin, tucusituzumab, umavizumab,urtoxazumab, and visilizumab.

A “metabolite” is a product produced through metabolism in the body of aspecified compound or salt thereof. Metabolites of a compound may beidentified using routine techniques known in the art and theiractivities determined using tests such as those described herein. Suchproducts may result e.g. from the oxidation, reduction, hydrolysis,amidation, deamidation, esterification, deesterification, enzymaticcleavage, and the like, of the administered compound. Accordingly, theinvention includes metabolites of compounds of the invention, includingcompounds produced by a process comprising contacting a compound of thisinvention with a mammal for a period of time sufficient to yield ametabolic product thereof.

The term “package insert” is used to refer to instructions customarilyincluded in commercial packages of therapeutic products, that containinformation about the indications, usage, dosage, administration,contraindications and/or warnings concerning the use of such therapeuticproducts.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “stereoisomers” refers to compounds which have identicalchemical constitution, but differ with regard to the arrangement of theatoms or groups in space.

“Diastereomer” refers to a stereoisomer with two or more centers ofchirality and whose molecules are not mirror images of one another.Diastereomers have different physical properties, e.g. melting points,boiling points, spectral properties, and reactivities. Mixtures ofdiastereomers may separate under high resolution analytical proceduressuch as electrophoresis and chromatography.

“Enantiomers” refer to two stereoisomers of a compound which arenon-superimposable mirror images of one another.

Stereochemical definitions and conventions used herein generally followS. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S.,“Stereochemistry of Organic Compounds”, John Wiley & Sons, Inc., NewYork, 1994. The compounds of the invention may contain asymmetric orchiral centers, and therefore exist in different stereoisomeric forms.It is intended that all stereoisomeric forms of the compounds of theinvention, including but not limited to, diastereomers, enantiomers andatropisomers, as well as mixtures thereof such as racemic mixtures, formpart of the present invention. Many organic compounds exist in opticallyactive forms, i.e., they have the ability to rotate the plane ofplane-polarized light. In describing an optically active compound, theprefixes D and L, or R and S, are used to denote the absoluteconfiguration of the molecule about its chiral center(s). The prefixes dand l or (+) and (−) are employed to designate the sign of rotation ofplane-polarized light by the compound, with (−) or l meaning that thecompound is levorotatory. A compound prefixed with (+) or d isdextrorotatory. For a given chemical structure, these stereoisomers areidentical except that they are mirror images of one another. A specificstereoisomer may also be referred to as an enantiomer, and a mixture ofsuch isomers is often called an enantiomeric mixture. A 50:50 mixture ofenantiomers is referred to as a racemic mixture or a racemate, which mayoccur where there has been no stereoselection or stereospecificity in achemical reaction or process. The terms “racemic mixture” and “racemate”refer to an equimolar mixture of two enantiomeric species, devoid ofoptical activity.

The term “tautomer” or “tautomeric form” refers to structural isomers ofdifferent energies which are interconvertible via a low energy barrier.For example, proton tautomers (also known as prototropic tautomers)include interconversions via migration of a proton, such as keto-enoland imine-enamine isomerizations. Valence tautomers includeinterconversions by reorganization of some of the bonding electrons.

The phrase “pharmaceutically acceptable salt” as used herein, refers topharmaceutically acceptable organic or inorganic salts of a compound ofthe invention. Exemplary salts include, but are not limited, to sulfate,citrate, acetate, oxalate, chloride, bromide, iodide, nitrate,bisulfate, phosphate, acid phosphate, isonicotinate, lactate,salicylate, acid citrate, tartrate, olcate, tannate, pantothenate,bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate,gluconate, glucuronate, saccharate, formate, benzoate, glutamate,methanesulfonate “mesylate”, ethanesulfonate, benzenesulfonate,p-toluenesulfonate, and pamoate (i.e.,1,1′-methylene-bis(2-hydroxy-3-naphthoate)) salts. A pharmaceuticallyacceptable salt may involve the inclusion of another molecule such as anacetate ion, a succinate ion or other counter ion. The counter ion maybe any organic or inorganic moiety that stabilizes the charge on theparent compound. Furthermore, a pharmaceutically acceptable salt mayhave more than one charged atom in its structure. Instances wheremultiple charged atoms are part of the pharmaceutically acceptable saltcan have multiple counter ions. Hence, a pharmaceutically acceptablesalt can have one or more charged atoms and/or one or more counter ion.

If the compound of the invention is a base, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method available in theart, e.g., treatment of the free base with an inorganic acid, such ashydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,methanesulfonic acid, phosphoric acid and the like, or with an organicacid, such as acetic acid, trifluoroacetic acid, maleic acid, succinicacid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalicacid, glycolic acid, salicylic acid, a pyranosidyl acid, such asglucuronic acid or galacturonic acid, an alpha hydroxy acid, such ascitric acid or tartaric acid, an amino acid, such as aspartic acid orglutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid,a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid,or the like.

If the compound of the invention is an acid, the desiredpharmaceutically acceptable salt may be prepared by any suitable method,e.g., treatment of the free acid with an inorganic or organic base, suchas an amine (primary, secondary or tertiary), an alkali metal hydroxideor alkaline earth metal hydroxide, or the like. Illustrative examples ofsuitable salts include, but are not limited to, organic salts derivedfrom amino acids, such as glycine and arginine, ammonia, primary,secondary, and tertiary amines, and cyclic amines, such as piperidine,morpholine and piperazine, and inorganic salts derived from sodium,calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminumand lithium.

The phrase “pharmaceutically acceptable” indicates that the substance orcomposition must be compatible chemically and/or toxicologically, withthe other ingredients comprising a formulation, and/or the mammal beingtreated therewith.

A “solvate” refers to an association or complex of one or more solventmolecules and a compound of the invention. Examples of solvents thatform solvates include, but are not limited to, water, isopropanol,ethanol, methanol, DMSO, ethylacetate, acetic acid, and ethanolamine.

The terms “compound of this invention,” and “compounds of the presentinvention” and “compounds of Formula I” include compounds of Formulas Iand stereoisomers, geometric isomers, tautomers, solvates, metabolites,and pharmaceutically acceptable salts and prodrugs thereof.

Any formula or structure given herein, including Formula I compounds, isalso intended to represent hydrates, solvates, and polymorphs of suchcompounds, and mixtures thereof.

Formula I Tricyclic PI3K Inhibitor Compounds

The present invention provides tricyclic PI3K inhibitor compounds ofFormula I, and pharmaceutical formulations thereof, which arepotentially useful in the treatment of diseases, conditions and/ordisorders modulated by PI3 kinases. More specifically, the presentinvention provides compounds of Formula I:

and stereoisomers, geometric isomers, tautomers, and pharmaceuticallyacceptable salts thereof, wherein:

the dashed lines indicate an optional double bond, and at least onedashed line is a double bond;

X¹ is S, O, N, NR^(a), CR¹, C(R¹)₂, or —C(R¹)₂O—;

X² is C, CR² or N;

X³ is C, CR³ or N;

A is a 5, 6, or 7-membered carbocyclyl or heterocyclyl ring fused to X²and X³, optionally substituted with one or more R⁵ groups;

R^(a) is H, C₁-C₁₂ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₁₂alkylene)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀heterocyclyl), —(C₁-C₁₂ alkylene)-C(═O)—(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂alkylene)-(C₆-C₂₀ aryl), and —(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl),where alkyl, alkenyl, alkynyl, alkylene, carbocyclyl, heterocyclyl,aryl, and heteroaryl are optionally substituted with one or more groupsindependently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —C(CH₃)₃,—CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH, —CH₂OCH₃, —CN, —CF₃, —CO₂H, —COCH₃,—COC(CH₃)₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —C(CH₃)₂CONH₂, —NO₂,—NH₂, —NHCH₃, —N(CH₃)₂, —NHCOCH₃, —NHS(O)₂CH₃, —N(CH₃)C(CH₃)₂CONH₂,—N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —S(O)₂CH,cyclopropyl, cyclobutyl, oxetanyl, morpholino, and1,1-dioxo-thiopyran-4-yl;

R¹, R², and R³ are independently selected from H, F, Cl, Br, I, —CH₃,—CH₂CH₃, —C(CH₃)₃, —CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH, —CH₂OCH₃, —CN, —CF₃,—CO₂H, —COCH₃, —COC(CH)₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂,—C(CH₃)₂CONH₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCOCH₃, —NHS(O)₂CH₃,—N(CH₃)C(CH₃)₂CONH₂, —N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃,—S(O)₂N(CH₃)₂, —SCH₃, —S(O)₂CH₃, cyclopropyl, cyclobutyl, oxetanyl,morpholino, and 1,1-dioxo-thiopyran-4-yl;

R⁴ is selected from C₆-C₂₀ aryl, C₂-C₂₀ heterocyclyl and C₁-C₂₀heteroaryl, each of which are optionally substituted with one or moregroups R⁶ groups independently selected from F, Cl, Br, I, —CH₃,—CH₂CH₃, —CH(CH₃)₂, —CH₂CH(CH₃)₂, —CH₂CH₃, —CH₂CN, —CN, —CF₃, —CH₂OH,—CO₂H, —CONH₂, —CONH(CH₃), —CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —NHCOCH₃,—OH, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —SH, —NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH₃,—NHS(O)₂CH₃, —N(CH₃)C(═O)OC(CH₃)₃, —S(O)₂CH₃, benzyl, benzyloxy,morpholinyl, morpholinomethyl, and 4-methylpiperazin-1-yl; and

R⁵ is independently selected from C₁-C₁₂ alkyl, C₂-C₈ alkenyl, C₂-C₈alkynyl, —(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂alkylene)-(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-C(═O)—(C₂-C₂₀heterocyclyl), —(C₁-C₁₂ alkylene)-(C₆-C₂₀ aryl), and —(C₁-C₁₂alkylene)-(C₁-C₂₀ heteroaryl); or two geminal R⁵ groups form a 3, 4, 5,or 6-membered carbocyclyl or heterocyclyl ring, where alkyl, alkenyl,alkynyl, alkylene, carbocyclyl, heterocyclyl, aryl, and heteroaryl areoptionally substituted with one or more groups independently selectedfrom F, Cl, Br, I, —CH₃, —CH₂CH₃, —C(CH₃)₂, —CH₂OH, —CH₂CH₂OH,—C(CH)₂OH, —CH₂OCH₃, —CN, —CH₂F, —CHF₂, —CF₃, —CO₂H, —COCH₃, —COC(CHO)₃,—CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —C(CH₃)₂CONH₂, —NO₂, —NH₂,—NHCH₃, —N(CH₃)₂, —NHCOCH₃, —NHS(O)₂CH₃, —N(CH₃)C(CH₃)₂CONH₂,—N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —S(O)₂CH₃,cyclopropyl, cyclobutyl, oxetanyl, morpholino, and1,1-dioxo-thiopyran-4-yl;

mor is selected from:

optionally substituted with one or more R⁷ groups independently selectedfrom F, Cl, Br, I, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃,—CH₂OCH₃, —CHF₂, —CN, —CF₃, —CH₂OH, —CH₂OCH₃, —CH₂CH₂OH, —CH₂C(CH₃)₂OH,—CH(CH₃)OH, —CH(CH₂CH₃)OH, —CH₂CH(OH)CH₃, —C(CH₃)₂OH, —C(CH₃)₂OCH₃,—CH(CH₃)F, —C(CH₃)F₂, —CH(CH₂CH₃)F, —C(CH₂CH)₂F, —CO₂H, —CONH₂,—CON(CH₂CH₃)₂, —COCH₃, —CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂,—NHCH₂CH₃, —NHCH(CH₃)₂, —NHCH₂CH₂OH, —NHCH₂CH₂OCH₃, —NHCOCH₃,—NHCOCH₂CH₃, —NHCOCH₂OH, —NHS(O)₂CH₃, —N(CH₃)S(O)₂CH₃, ═O, —OH, —OCH₃,—OCH₂CH₃, —OCH(CH₃)₂, —SH, —NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH₃, —S(O)CH₃,—S(O)CH₂CH₃, —S(O)₂CH₃, —S(O)₂NH₂, —S(O)₂NHCH₃, —S(O)₂N(CH₃)₂, and—CH₂S(O)₂CH₃.

Further it is to be understood that every embodiment relating to aspecific residue X¹, X², X³, A, R^(a), R¹, R², R³, R⁴, R⁵, R⁶, R⁷ andmor as disclosed herein may be combined with any other embodimentrelating to another residue X¹, X², X³, A, R^(a), R¹, R², R³, R⁴, R⁵,R⁶, R⁷ and mor as disclosed herein.

Exemplary embodiments of compounds of the invention include FormulasIa-n:

Exemplary embodiments of Formula I compounds include:

where the A heterocyclyl ring is optionally substituted with one or moreR⁵ groups.

Exemplary Formula I compounds include:

where the A heterocyclyl ring is optionally substituted with one or moreR⁵ groups.

In exemplary Formula I compounds, the A heterocyclyl ring is optionallysubstituted with one, two or three R⁵ groups.

In exemplary Formula I compounds, the A heterocyclyl ring is optionallysubstituted with two R⁵ groups.

Exemplary Formula I compounds include two geminal R⁵ groups which formcyclopropyl, cyclobutyl, cyclopentyl, tetrahydrofuryl,tetrahydropyranyl, oxetanyl, azetidinyl, pyrrolidinyl, piperidyl,piperazinyl, cyclohexyl, morpholino, or 1,1-dioxo-thiopyran-4-yl.

Exemplary Formula I compounds include wherein R⁴ is phenyl substitutedwith one or more groups selected from F, Cl, Br, I, —CH₃, —CH₂CH₃,—CH(CH₃)₂, —CN, —CF₃, —CH₂OH, —CO₂H, —CONH₂, —CONH(CH₃), —CON(CH₃)₂,—NO₂, —NH₂, —NHCH, —NHCOCH₃, —OH, —OCH₃, —OCH₂CH₃, —OCH(CH)₂, —SH,—NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH₃, —NHS(O)₂CH₃, —N(CH₃)C(═O)OC(CH₃)₃, and—S(O)₂CH₃.

Exemplary Formula I compounds include wherein R⁴ is an optionallysubstituted bicyclic heteroaryl group selected from 1H-indazole,1H-indole, indolin-2-one, 1-(indolin-1-yl)ethanone,1H-benzo[d][1,2,3]triazole, 1H-pyrazolo[3,4-b]pyridine,1H-pyrazolo[3,4-d]pyrimidine, 1H-benzo[d]imidazole,1H-benzo[d]imidazol-2(3H)-one, 1H-pyrazolo[3,4-e]pyridine,1H-pyrrolo[2,3-e]pyridine, 3H-imidazo[4,5-e]pyridine,7H-pyrrolo[2,3-d]pyrimidine, 7H-purine, 1H-pyrazolo[4,3-d]pyrimidine,5H-pyrrolo[3,2-d]pyrimidine, 2-amino-1H-purin-6(9H)-one, quinoline,quinazoline, quinoxaline, isoquinoline, isoquinolin-1(2H)-one,3,4-dihydroisoquinolin-1(2H)-one, 3,4-dihydroquinolin-2(1H)-one,quinazolin-2(1H)-one, quinoxalin-2(1H)-one, 1,8-naphthyridine,pyrido[3,4-d]pyrimidine, and pyrido[3,2-b]pyrazine.

Exemplary Formula I compounds include wherein optionally substituted R⁴is selected from:

where the wavy line indicates the site of attachment.Exemplary Formula I compounds include wherein optionally substituted R⁴is selected from:

where the wavy line indicates the site of attachment.

In one embodiment of the invention R⁴ is 1H-indazol-4-yl.

Exemplary Formula I compounds include wherein R⁴ is an optionallysubstituted monocyclic heteroaryl group selected from 2-furanyl,3-furanyl, 2-imidazolyl, 4-imidazolyl, 3-isoxazolyl, 4-isoxazolyl,5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-pyrazolyl,4-pyrazolyl, 2-pyrazinyl, 3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl,2-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-pyridyl, 3-pyridyl,4-pyridyl, 2-pyrrolyl, 3-pyrrolyl, 2-thienyl, 3-thienyl, 5-tetrazolyl,1-tetrazolyl, 2-tetrazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,3-triazolyl, and 1-triazolyl.

Exemplary Formula I compounds include optionally substituted R⁴ isselected from:

where the wavy line indicates the site of attachment.

Exemplary Formula I compounds include wherein R⁴ is an optionallysubstituted monocyclic heteroaryl group selected from pyridyl,pyrimidinyl or pyrazolyl.

Exemplary Formula I compounds include wherein R⁴ is an optionallysubstituted pyrimidinyl.

In one embodiment of the invention R⁴ is 2-aminopyrimidin-5-yl.

Exemplary Formula I compounds include optionally substituted R⁴:

where the wavy line indicates the site of attachment.

Exemplary Formula I compounds include mor:

where the wavy line indicates the site of attachment, optionallysubstituted with one or more R⁷ groups independently selected from F,Cl, Br, I, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, —CH₂OCH₃,—CHF₂, —CN, —CF₃, —CH₂OH, —CH₂OCH₃, —CH₂CH₂OH, —CH₂C(CH₃)₂OH,—CH(CH₃)OH, —CH(CH₂CH₃)OH, —CH₂CH(OH)CH₃, —C(CH₃)₂OH, —C(CH₃)₂OCH₃,—CH(CH₃)F, —C(CH₃)F₂, —CH(CH₂CH₃)F, —C(CH₂CH₃)₂F, —CO₂H, —CONH₂,—CON(CH₂CH₃)₂, —COCH₃, —CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —N(CH)₂,—NHCH₂CH₃, —NHCH(CH₃)₂, —NHCH₂CH₂OH, —NHCH₂CH₂OCH₃, —NHCOCH₃,—NHCOCH₂CH₃, —NHCOCH₂OH, —NHS(O)₂CH₃, —N(CH₃)S(O)₂CH₃, ═O, —OH, —OCH₃,—OCH₂CH₃, —OCH(CH₃)₂, —SH, —NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH, —S(O)CH₃,—S(O)CH₂CH₃, —S(O)₂CH₃, —S(O)₂NH₂, —S(O)₂NHCH₃, —S(O)₂N(CH₃)₂, and—CH₂S(O)₂CH₃.

In one embodiment of the invention, one or more R⁵ groups are C₁-C₁₂alkyl optionally substituted with one or more groups selected from F,Cl, Br, I, —CH₃, —CH₂CH₃, —C(CH₃)₃, —CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH,—CH₂OCH₃, —CN, —CH₂F, —CHF₂, —CF₃, —CO₂H, —COCH₃, —COC(CH₃)₃, —CO₂CH₃,—CONH₂, —CONHCH₃, —CON(CH₃)₂, —C(CH₃)₂CONH₂, —NO₂, —NH₂, —NHCH₃,—N(CH₃)₂, —NHCOCH₃, —NHS(O)₂CH₃, —N(CH₃)C(CH₃)₂CONH₂,—N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —S(O)₂CH₃,cyclopropyl, cyclobutyl, oxetanyl, morpholino, and1,1-dioxo-thiopyran-4-yl.

In one embodiment of the invention, R⁵ is methyl optionally substitutedwith one or more groups as defined herein. In one embodiment suchsubstituents are F, OH and ═O.

In one embodiment of the invention one or more R⁵ groups areindependently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —C(CH₃)₃,—CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH, —CH₂OCH₃, —CN, —CH₂F, —CHF₂, —CF₃, —CO₂H,—COCH₃, —COC(CH₃)₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂,—C(CH₃)₂CONH₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCOCH₃, —NHS(O)₂CH₃,—N(CH₃)C(CH₃)₂CONH₂, —N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃,—S(O)₂N(CH₃)₂, —SCH₃, —S(O)₂CH₃, cyclopropyl, cyclobutyl, oxetanyl,morpholino, and 1,1-dioxo-thiopyran-4-yl.

The Formula I compounds of the invention may contain asymmetric orchiral centers, and therefore exist in different stereoisomeric forms.It is intended that all stereoisomeric forms of the compounds of theinvention, including but not limited to, diastereomers, enantiomers andatropisomers, as well as mixtures thereof such as racemic mixtures, formpart of the present invention.

In addition, the present invention embraces all geometric and positionalisomers. For example, if a Formula I compound incorporates a double bondor a fused ring, the cis- and trans-forms, as well as mixtures thereof,are embraced within the scope of the invention. Both the singlepositional isomers and mixture of positional isomers are also within thescope of the present invention.

In the structures shown herein, where the stereochemistry of anyparticular chiral atom is not specified, then all stereoisomers arecontemplated and included as the compounds of the invention. Wherestereochemistry is specified by a solid wedge or dashed linerepresenting a particular configuration, then that stereoisomer is sospecified and defined.

The compounds of the present invention may exist in unsolvated as wellas solvated forms with pharmaceutically acceptable solvents such aswater, ethanol, and the like, and it is intended that the inventionembrace both solvated and unsolvated forms.

The compounds of the present invention may also exist in differenttautomeric forms, and all such forms are embraced within the scope ofthe invention. The term “tautomer” or “tautomeric form” refers tostructural isomers of different energies which are interconvertible viaa low energy barrier. For example, proton tautomers (also known asprototropic tautomers) include interconversions via migration of aproton, such as keto-enol and imine-enamine isomerizations. Valencetautomers include interconversions by reorganization of some of thebonding electrons.

The present invention also embraces isotopically-labeled compounds ofthe present invention which are identical to those recited herein, butfor the fact that one or more atoms are replaced by an atom having anatomic mass or mass number different from the atomic mass or mass numberusually found in nature. All isotopes of any particular atom or elementas specified are contemplated within the scope of the compounds of theinvention, and their uses. Exemplary isotopes that can be incorporatedinto compounds of the invention include isotopes of hydrogen, carbon,nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine and iodine,such as ²H(D), ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³²P, ³³P,³⁵S, ¹⁸F, ³⁶Cl, ¹²³I and ¹²⁵I. Certain isotopically-labeled compounds ofthe present invention (e.g., those labeled with ³H and ¹⁴C) are usefulin compound and/or substrate tissue distribution assays. Tritiated (³H)and carbon-14 (¹⁴C) isotopes are useful for their ease of preparationand detectability. Further, substitution with heavier isotopes such asdeuterium (i.e., ²H) may afford certain therapeutic advantages resultingfrom greater metabolic stability (e.g., increased in vivo half-life orreduced dosage requirements) and hence may be preferred in somecircumstances. Positron emitting isotopes such as ¹⁵O, ¹³N, ¹¹C and ¹⁸Fare useful for positron emission tomography (PET) studies to examinesubstrate receptor occupancy. Isotopically labeled compounds of thepresent invention can generally be prepared by following proceduresanalogous to those disclosed in the Schemes and/or in the Examplesherein below, by substituting an isotopically labeled reagent for anon-isotopically labeled reagent.

Biological Evaluation

Determination of the PI3 kinase activity of a Formula I compound ispossible by a number of direct and indirect detection methods. Certainexemplary compounds described herein were assayed for their p110α(alpha), and other isoform, PI3K binding activity (Example 901) and invitro activity against tumor cells (Example 902). Certain exemplarycompounds of the invention had PI3K binding activity IC₅₀ values lessthan 10 nM. Certain compounds of the invention had tumor cell-basedactivity EC₅₀ values less than 100 nM.

The cytotoxic or cytostatic activity of Formula I exemplary compoundswas measured by: establishing a proliferating mammalian tumor cell linein a cell culture medium, adding a Formula I compound, culturing thecells for a period from about 6 hours to about 5 days; and measuringcell viability (Example 902). Cell-based in vitro assays were used tomeasure viability, i.e. proliferation (IC₅₀), cytotoxicity (EC₅₀), andinduction of apoptosis (caspase activation).

The in vitro potency of Formula I exemplary compounds was measured bythe cell proliferation assay, CellTiter-Glo® Luminescent Cell ViabilityAssay, commercially available from Promega Corp., Madison, Wis. (Example902). This homogeneous assay method is based on the recombinantexpression of Coleoptera luciferase (U.S. Pat. Nos. 5,583,024;5,674,713; 5,700,670) and determines the number of viable cells inculture based on quantitation of the ATP present, an indicator ofmetabolically active cells (Crouch et al (1993) J. Immunol. Meth.160:81-88: U.S. Pat. No. 6,602,677). The CellTiter-Glo® Assay wasconducted in 96 or 384 well format, making it amenable to automatedhigh-throughput screening (HTS) (Cree et al (1995) AntiCancer Drugs6:398-404). The homogeneous assay procedure involves adding the singlereagent (CellTiter-Glo® Reagent) directly to cells cultured inserum-supplemented medium. Cell washing, removal of medium and multiplepipetting steps are not required. The system detects as few as 15cells/well in a 384-well format in 10 minutes after adding reagent andmixing.

The homogeneous “add-mix-measure” format results in cell lysis andgeneration of a luminescent signal proportional to the amount of ATPpresent. The amount of ATP is directly proportional to the number ofcells present in culture. The CellTiter-Glo® Assay generates a“glow-type” luminescent signal, produced by the luciferase reaction,which has a half-life generally greater than five hours, depending oncell type and medium used. Viable cells are reflected in relativeluminescence units (RLU). The substrate, Beetle Luciferin, isoxidatively decarboxylated by recombinant firefly luciferase withconcomitant conversion of ATP to AMP and generation of photons. Theextended half-life eliminates the need to use reagent injectors andprovides flexibility for continuous or batch mode processing of multipleplates. This cell proliferation assay can be used with various multiwellformats, e.g. 96 or 384 well format. Data can be recorded by luminometeror CCD camera imaging device. The luminescence output is presented asrelative light units (RLU), measured over time.

The anti-proliferative effects of Formula I exemplary compounds weremeasured by the CellTiter-Glo® Assay (Example 902) against several tumorcell lines. Potency EC₅₀ values were established for the testedcompounds. The range of in vitro cell potency activities was about 100nM to about 10 μM. Certain tested compounds had EC₅₀ values of less than1 micromolar (1 μM) in stopping proliferation of certain tumor celllines.

Certain ADME properties were measured for certain exemplary compounds byassays including: Caco-2 Permeability (Example 903), HepatocyteClearance (Example 904), Cytochrome P450 Inhibition (Example 905),Cytochrome P450 Induction (Example 906), Plasma Protein Binding (Example907), and hERG channel blockage (Example 908).

Certain exemplary compounds were tested for efficacy in dose escalationstudies in tumor-bearing Taconic NCR nude mouse models (Example 909).The U-87 MG Merchant (an in-house variant derived from U-87 MG cellsfrom ATCC, Manassas, Va.) subcutaneous xenograft mouse model wasemployed to test Formula I compounds at escalating doses along withVehicle (MCT, negative control). Tumor growth delay was measuredfollowing once daily oral dosing for <28 days. Body weight change overthe course of treatment was measured as an indicator of safety. Thedose- and time-dependent pharmacokinetic and pharmacodynamic response ofdrug administration in this same subcutaneous tumor xenograft model wasalso examined (Example 913).

Blood-brain barrier penetrant [properties] potential was assessed invitro using MDCK cells stably transfected with P-glycoprotein (MDR1) orbcrp1 (Example 911). Brain penetration was determined in vivo bymeasuring compound concentrations (Example 912) and/or by measuring themodulation of the PI3K pathway (Example 913) in the brain of micefollowing a single IV or oral dose. Brain tumor efficacy was measured inExample 914 by GS-2 (human glioblastoma muliforme (GBM) engineered toexpress luciferase). The effect of once daily oral dosing on the growthof GS-2 intracranial implants was evaluated by magnetic resonanceimaging (MRI). Mice with tumor xenografts of U-87 MG cells were dosedwith drug or vehicle and samples were analyzed for PK, PD, and/or IHCanalysis (Example 915).

Exemplary Formula I compounds No. 101-177 in Table I were made,characterized, and tested for inhibition of PI3K alpha (IC₅₀ or K_(i)binding to p110 alpha less than 1 micromolar, μM) and selectivityaccording to the methods of this invention, and have the followingstructures and corresponding names (ChemBioDraw Ultra, Version 11.0,CambridgeSoft Corp., Cambridge Mass.).

TABLE 1 IC₅₀, Ki (μmole) P13K, p110 alpha binding No. Structure Nameassay 101

1-[4-(3a,8-dimethyl-7- morpholin-4-yl-3,3a,8,8a-tetrahydro-2h-1-oxa-4,6,8- triaza-cyclopenta[a]inden-5-yl)-phenyl]-3-ethyl-urea 2.1 102

5-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)- 4-methylpyrimidin-2-amine 0.0018 103

5-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2- yl)pyrimidin-2-amine 0.00209 104

5-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)- 4-(trifluoromethyl)pyridyl-2- amine0.00389 105

5-(4-morpholino-8,9-dihydro- 7h-[1,3]oxazino[2,3-e]purin-2-yl)pyrimidin-2-amine 0.00684 106

5-(4-morpholino-6,7,8,9- tetrahydropyrido[2,1-e]purin-2-yl)pyrimidin-2-amine 0.00388 107

5-(4-morpholino-6,7,8,9- tetrahydropyrido[2,1-e]purin-2-yl)pyridin-2-amine 0.0507 108

5-(4-morpholino-8,9-dihydro- 6h-[1,4]oxazino[3,4-e]purin-2-yl)-4-(trifluoromethyl)pyridyl-2- amine 0.0112 109

5-(4-morpholino-7,8-dihydro- 6h-pyrrolo[2,1-e]purin-2-yl)pyrimidin-2-amine 0.00826 110

6,6-dimethyl-4-morpholino-2- (1H-pyrrolo[2,3-b]pyridin-5-yl)-8,9-dihydro-6H- [1,4]oxazino[3,4-e]purine 0.0140 111

5-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyridin-2-amine 0.0186 112

5-(4-morpholino-8,9- dihydrospiro[[1,3]oxazino[2,3-e]purine-7,1′-cyclopropane]-2- yl)pyrimidin-2-amine 0.00137 113

5-(4-morpholino-8,9-dihydro- 6h-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine 0.00432 114

5-(4-morpholino-8,9- dihydrospiro[[1,4]oxazino[3,4-e]purine-6,3′-oxetane]-2- yl)pyrimidin-2-amine 0.00245 115

5-(7,7-dimethyl-4-morpholino- 8,9-dihydro-7h-[1,3]oxazino[2,3-e]purin-2- yl)pyrimidin-2-amine 0.00509 116

5-(4-morpholino-6- (trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyridin-2-amine 0.0452 117

5-(6,6-(hexadeuterio)dimethyl- 4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2- yl)pyrimidin-2-amine 0.00259 118

(S)-5-(6-ethyl-6-methyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyrimidin-2-amine 0.00311 119

5-(6,6,9-trimethyl-4- morpholino-6h- [1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine 0.00473 120

(R)-5-(6-ethyl-6-methyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyrimidin-2-amine 0.00596 121

5-(1-morpholin-4-yl-5,6,8a,9- tetrahydro-8h-7,10-dioxa-2,4,4b-triaza-phenanthren-3-yl)- pyrimidin-2-ylamine 0.00572 122

5-((S)-6-Morpholin-4-yl- 2,3,3a,4-tetrahydro-1H-5-oxa- 7,9,9b-triaza-cyclopenta[a]naphthalen-8-yl)- pyrimidin-2-ylamine 0.0779 123

4-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)aniline 0.541 124

1-(4-(6,6-dimethyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenyl)-3-methylurea 0.00671 125

6,6-dimethyl-4-morpholino-2- (1H-pyrazol-4-yl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine 0.382 126

4-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyridin-2-amine 0.685 127

6,6-dimethyl-2-(1-methyl-1H- pyrazol-4-yl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 128

3-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenol 0.00458 129

2-(1H-indazol-5-yl)-6,6- dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 130

6,6-dimethyl-2-(2-(4- methylpiperazin-1-yl) yridine-4-yl)-4-morpholino-8,9-dihydro- 6H-[1,4]oxazino[3,4-e]purine >0.695 131

N-(2-(6,6-dimethyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenyl)methanesulfonamide 0.638 132

6,6-dimethyl-4-morpholino-2- (6-morpholinopyridin-3-yl)-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 133

2-(1-benzyl-1H-pyrazol-4-yl)- 6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 134

2-(2-isopropoxypyridin-3-yl)- 6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 135

N-(2-(6,6-dimethyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenyl)acetamide >0.695 136

2-(3,5-dimethyl-1H-pyrazol-4- yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H- [1,4]oxazino[3,4-e]purine >0.695 137

5-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyridine-2-ol >0.695 138

6-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyridine-3-amine >0.695 139

(R)-5-(4-morpholino-6- (trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyrimidin-2-amine 0.00357 140

(S)-5-(4-morpholino-6- (trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyrimidin-2-amine 0.00345 141

2-(1-ethyl-1H-pyrazol-4-yl)-6,6- dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 142

4-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)- N,N-dimethylbenzamide >0.695 143

tert-butyl 4-(6,6-dimethyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenyl(methyl)carbamate >0.695 144

2-(3-(6,6-dimethyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenyl)acetonitrile >0.695 145

6,6-dimethyl-4-morpholino-2- (3-morpholinophenyl)-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 146

6,6-dimethyl-4-morpholino-2- (3-(morpholinomethyl)phenyl)-8,9-dihydro-6H- [1,4]oxazino[3,4-e]purine >0.695 147

2-(3-(benzyloxy)phenyl)-6,6- dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 148

2-(1-isobutyl-1H-pyrazol-4-yl)- 6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 149

6,6-dimethyl-2-(6-(4- methylpiperazin-1-yl) yridine-3-yl)-4-morpholino-8,9-dihydro- 6H-[1,4]oxazino[3,4-e]purine >0.695 150

2-(1H-indazol-4-yl)-6,6- dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine 0.0198 151

4-(6,6-ditnethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)benzonitrile >0.695 152

5-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)nicotinamide 0.0352 153

5-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)- N-methylpicolinamide >0.695 154

2-(4-(benzyloxy)phenyl)-6,6- dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 155

3-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)- N,N-dimethylaniline >0.695 156

6,6-dimethyl-2-(4-(4- methylpiperazin-1-yl)phenyl)-4-morpholino-8,9-dihydro-6H- [1,4]oxazino[3,4-e]purine >0.695 157

6,6-dimethyl-4-morpholino-2- (4-(piperidin-1-yl)phenyl)-8,9-dihydro-6H-[1,4]oxazino[3,4- e]purine >0.695 158

N-(5-(6,6-dimethyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyridine-2-yl)acetamide 0.0490 159

5-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)picolinamide — 160

6-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-c]purin-2- yl)pyridine-3-ol --- 161

(4-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenyl)(4-methylpiperazin-1-yl)methanone >0.695 162

N-cyclopropyl-3-(6,6-dimethyl- 4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)benzamide >0.695 163

5-(6,6-dimethyl-4-morpholino- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)- N,N-dimethylpyrazin-2-amine >0.695 164

1-(4-(6,6-dimethyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenyl)-3-ethylurea 0.0417 165

1-(4-(6,6-dimethyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenyl)-3-isopropylurea 0.337 166

(2-(2-aminopyrimidin-5-yl)-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine-6,6- diyl)dimethanol 0.0128 167

2-(2-aminopyrimidin-5-yl)-7- methyl-4-morpholino-8,9-dihydropyrazino[2,1-e]purin- 6(7H)-one 0.0215 168

5-(8,8-Dimethyl-1-morpholin-4- yl-5,8-dihydro-6H-7-oxa-9-thia-2,4-diaza-fluoren-3-yl)- pyrimidin-2-ylamine 0.00100 169

2-(1H-indazol-4-yl)-4- morpholino-6-(trifluoromethyl)- 8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine 0.0452 170

3-(4-morpholino-6- (trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)phenol 0.0797 171

5-(4-((2S,6R)-2,6- dimethylmorpholino)-6,6- dimethyl-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyrimidin-2-amine 0.01398 172

5-(4-(2,2-dimethylmorpholino)- 6,6-dimethyl-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyrimidin-2-amine 0.201 173

N-(5-(6,6-dimethyl-4- morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2- yl)pyrimidin-2-yl)acetamide 0.242 174

5-(4-((1S,4S)-2-oxa-5- azabicyclo[2.2.1]heptan-5-yl)-6,6-dimethyl-8,9-dihydro-6H- [1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine 0.394 175

2-(2-aminopyrimidin-5-yl)-6- methyl-4-morpholino-6,7-dihydropyrazino[2,1-e]purin- 8(9H)-one 0.00428 176

5-(6,7-dimethyl-4-morpholino- 6,7,8,9-tetrahydropyrazino[2,1-e]purin-2-yl)pyrimidin-2-amine 0.00689 177

5-(8,8-Dimethyl-1-morpholin-4- yl-5,6-dihydro-8H-7-oxa-2,4,4b-triaza-fluoren-3-yl)-pyrimidin-2- ylamine 0.000831

Administration of Formula I Compounds

The Formula I compounds of the invention may be administered by anyroute appropriate to the condition to be treated. Suitable routesinclude oral, parenteral (including subcutaneous, intramuscular,intravenous, intraarterial, intradermal, intrathecal and epidural),transdermal, rectal, nasal, topical (including buccal and sublingual),vaginal, intraperitoneal, intrapulmonary and intranasal. For localimmunosuppressive treatment, the compounds may be administered byintralesional administration, including perfusing or otherwisecontacting the graft with the inhibitor before transplantation. It willbe appreciated that the preferred route may vary with e.g. the conditionof the recipient. Where the compound is administered orally, it may beformulated as a pill, capsule, tablet, etc. with a pharmaceuticallyacceptable carrier or excipient. Where the compound is administeredparenterally, it may be formulated with a pharmaceutically acceptableparenteral vehicle and in a unit dosage injectable form, as detailedbelow.

A dose to treat human patients may range from about 10 mg to about 1000mg of Formula I compound. A typical dose may be about 100 mg to about300 mg of the compound. A dose may be administered once a day (QID),twice per day (BID), or more frequently, depending on thepharmacokinetic and pharmacodynamic properties, including absorption,distribution, metabolism, and excretion of the particular compound. Inaddition, toxicity factors may influence the dosage and administrationregimen. When administered orally, the pill, capsule, or tablet may beingested daily or less frequently for a specified period of time. Theregimen may be repeated for a number of cycles of therapy.

Methods of Treatment with Formula I Compounds

Formula I compounds of the present invention are useful for treatinghyperproliferative diseases, conditions and/or disorders including, butnot limited to, those characterized by over expression of lipid kinases,e.g. PI3 kinase. Accordingly, an aspect of this invention includesmethods of treating or preventing diseases or conditions that can betreated or prevented by inhibiting lipid kinases, including PI3. In oneembodiment, the method comprises administering to a mammal in needthereof a therapeutically effective amount of a compound of Formula I,or a stereoisomer, geometric isomer, tautomer, or pharmaceuticallyacceptable salt thereof. In one embodiment, a human patient is treatedwith a compound of Formula I and a pharmaceutically acceptable carrier,adjuvant, or vehicle, wherein said compound of Formula I is present inan amount to detectably inhibit PI3 kinase activity.

One embodiment of the invention includes a method of treating cancer ina patient comprised of administering to said patient a therapeuticallyeffective amount of a compound of this invention wherein the cancer isbreast, ovary, cervix, prostate, testis, genitourinary tract, esophagus,larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma,lung, epidermoid carcinoma, large cell carcinoma, non-small cell lungcarcinoma (NSCLC), small cell carcinoma, lung adenocarcinoma, bone,colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma,undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma,sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidneycarcinoma, renal, pancreatic, myeloid disorders, lymphoma, hairy cells,buccal cavity, naso-pharyngeal, pharynx, lip, tongue, mouth, smallintestine, colon-rectum, large intestine, rectum, brain and centralnervous system, Hodgkin's or leukemia. In one embodiment, the cancer isa brain cancer.

In one embodiment of the invention, the method further comprisesadministering to the patient an additional therapeutic agent selectedfrom a chemotherapeutic agent, an anti-angiogenesis therapeutic agent,an anti-inflammatory agent, an immunomodulatory agent, a neurotropicfactor, an agent for treating cardiovascular disease, an agent fortreating liver disease, an anti-viral agent, an agent for treating blooddisorders, an agent for treating diabetes, and an agent for treatingimmunodeficiency disorders. In one embodiment of the invention theadditional therapeutic agent is bevacizumab.

Formula I compounds may also be useful for treating hyperproliferativediseases characterized by over expression of protein kinases such asthose encoded by PIM; the genes Pim-1, Pim-2, and Pim-3 (ProviralInsertion, Moloney) which are implicated in lymphoma and solid-tumordevelopment (Cuypers et al. (1984) Cell, vol. 37 (1) pp. 141-50; Seltenet al. (1985) EMBO J. vol. 4 (7) pp. 1793-8; van der Lugt et al. (1995)EMBO J. vol. 14 (11) pp. 2536-44; Mikkers et al. (2002) Nature Genetics,vol. 32 (1) pp. 153-9; van Lohuizen et al. (1991) Cell, vol. 65 (5) pp.737-52.

Cancers which can be treated according to the methods of this inventioninclude, but are not limited to, breast, ovary, cervix, prostate,testis, genitourinary tract, esophagus, larynx, glioblastoma,neuroblastoma, stomach, skin, keratoacanthoma, lung, epidermoidcarcinoma, large cell carcinoma, non-small cell lung carcinoma (NSCLC),small cell carcinoma, lung adenocarcinoma, bone, colon, adenoma,pancreas, adenocarcinoma, thyroid, follicular carcinoma,undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma,sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidneycarcinoma, myeloid disorders, lymphoid disorders, hairy cells, buccalcavity and pharynx (oral), lip, tongue, mouth, pharynx, small intestine,colon-rectum, large intestine, rectum, brain and central nervous system,Hodgkin's and leukemia.

Formula I compounds may be useful for in vitro, in situ, and in vivodiagnosis or treatment of mammalian cells, organisms, or associatedpathological conditions, such as systemic and local inflammation,immune-inflammatory diseases such as rheumatoid arthritis, immunesuppression, organ transplant rejection, allergies, ulcerative colitis,Crohn's disease, dermatitis, asthma, systemic lupus crythematosus,Sjögren's Syndrome, multiple sclerosis, scleroderma/systemic sclerosis,idiopathic thrombocytopenic purpura (ITP), anti-neutrophil cytoplasmicantibodies (ANCA) vasculitis, chronic obstructive pulmonary disease(COPD), psoriasis, and for general joint protective effects.

Formula I compounds may be useful for treating conditions of the brainand central nervous system which require transport across theblood-brain barrier. Certain Formula I compounds have favorablepenetrant properties across the blood-brain barrier for delivery to thebrain. Disorders of the brain which may be effectively treated withFormula I compounds include metastatic and primary brain tumors, such asglioblastoma and melanoma.

Formula I compounds may be useful for treating ocular disorders such aswet and dry Age-related Macular Degeneration (AMD) and retina edema, bylocalized delivery to the eye. Certain Formula I compounds havefavorable properties for delivery to, and uptake into, the eye. CertainFormula I compounds may enhance efficacy and extend duration of responsefor treatment of wet AMD in combination with ranibizumab (LUCENTIS®,Genentech, Inc.) and bevacizumab (AVASTIN®, Genentech, Inc.).

Another aspect of this invention provides a compound of this inventionfor use in the treatment of the diseases or conditions described hereinin a mammal, e.g., a human, suffering from such disease or condition.Also provided is the use of a compound of this invention in thepreparation of a medicament for the treatment of the diseases andconditions described herein in a warm-blooded animal, such as a mammal,e.g. a human, suffering from such disorder.

Pharmaceutical Formulations

In order to use a Formula I compound for the therapeutic treatment(including prophylactic treatment) of mammals including humans, it isnormally formulated in accordance with standard pharmaceutical practiceas a pharmaceutical composition. According to this aspect of theinvention there is provided a pharmaceutical composition comprising acompound of this invention in association with a pharmaceuticallyacceptable diluent or carrier.

One embodiment of the invention comprises a pharmaceutical compositioncomprised of a compound of this invention and a pharmaceuticallyacceptable carrier, glidant, diluent, or excipient.

One embodiment of the invention comprises a process for making apharmaceutical composition which comprises combining a compound of thisinvention with a pharmaceutically acceptable carrier.

One embodiment of the invention includes a pharmaceutical composition asdescribed to above further comprising an additional therapeutic agentselected from a chemotherapeutic agent, an anti-inflammatory agent, animmunomodulatory agent, a neurotropic factor, an agent for treatingcardiovascular disease, an agent for treating liver disease, ananti-viral agent, an agent for treating blood disorders, an agent fortreating diabetes, and an agent for treating immunodeficiency disorders.

A typical formulation is prepared by mixing a Formula I compound and acarrier, diluent or excipient. Suitable carriers, diluents andexcipients are well known to those skilled in the art and includematerials such as carbohydrates, waxes, water soluble and/or swellablepolymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents,water and the like.

The particular carrier, diluent or excipient used will depend upon themeans and purpose for which the compound of the present invention isbeing applied. Solvents are generally selected based on solventsrecognized by persons skilled in the art as safe (GRAS) to beadministered to a mammal. In general, safe solvents are non-toxicaqueous solvents such as water and other non-toxic solvents that aresoluble or miscible in water. Suitable aqueous solvents include water,ethanol, propylene glycol, polyethylene glycols (e.g., PEG 400, PEG300), etc. and mixtures thereof. The formulations may also include oneor more buffers, stabilizing agents, surfactants, wetting agents,lubricating agents, emulsifiers, suspending agents, preservatives,antioxidants, opaquing agents, glidants, processing aids, colorants,sweeteners, perfuming agents, flavoring agents and other known additivesto provide an elegant presentation of the drug (i.e., a compound of thepresent invention or pharmaceutical composition thereof) or aid in themanufacturing of the pharmaceutical product (i.e., medicament).

The formulations may be prepared using conventional dissolution andmixing procedures. For example, the bulk drug substance (i.e., compoundof the present invention or stabilized form of the Formula I compound(e.g., complex with a cyclodextrin derivative or other knowncomplexation agent) is dissolved in a suitable solvent in the presenceof one or more of the excipients described above. The compound of thepresent invention is typically formulated into pharmaceutical dosageforms to provide an easily controllable dosage of the drug and to enablepatient compliance with the prescribed regimen.

The pharmaceutical composition (or formulation) may be packaged in avariety of ways depending upon the method of administering the drug.Generally, an article for distribution includes a container havingdeposited therein the pharmaceutical formulation in an appropriate form.Suitable containers are well known to those skilled in the art andinclude materials such as bottles (plastic and glass), sachets,ampoules, plastic bags, metal cylinders, and the like. The container mayalso include a tamper-proof assemblage to prevent indiscreet access tothe contents of the package. In addition, the container has depositedthereon a label that describes the contents of the container. The labelmay also include appropriate warnings.

Pharmaceutical formulations of the compounds of the present inventionmay be prepared for various routes and types of administration. Forexample, a compound of Formula I having the desired degree of purity mayoptionally be mixed with pharmaceutically acceptable diluents, carriers,excipients or stabilizers (Remington's Pharmaceutical Sciences (1980)16^(th) edition, Osol, A. Ed.), in the form of a lyophilizedformulation, milled powder, or an aqueous solution. Formulation may beconducted by mixing at ambient temperature at the appropriate pH, and atthe desired degree of purity, with physiologically acceptable carriers,i.e., carriers that are non-toxic to recipients at the dosages andconcentrations employed. The pH of the formulation depends mainly on theparticular use and the concentration of compound, but may range fromabout 3 to about 8. Formulation in an acetate buffer at pH 5 is asuitable embodiment.

The compound of this invention for use herein is preferably sterile. Inparticular, formulations to be used for in vivo administration must besterile. Such sterilization is readily accomplished by filtrationthrough sterile filtration membranes.

The compound ordinarily can be stored as a solid composition, alyophilized formulation or as an aqueous solution.

The pharmaceutical compositions of the invention comprising a Formula Icompound will be formulated, dosed and administered in a fashion, i.e.,amounts, concentrations, schedules, course, vehicles and route ofadministration, consistent with good medical practice. Factors forconsideration in this context include the particular disorder beingtreated, the particular mammal being treated, the clinical condition ofthe individual patient, the cause of the disorder, the site of deliveryof the agent, the method of administration, the scheduling ofadministration, and other factors known to medical practitioners. The“therapeutically effective amount” of the compound to be administeredwill be governed by such considerations, and is the minimum amountnecessary to prevent, ameliorate, or treat the coagulation factormediated disorder. Such amount is preferably below the amount that istoxic to the host or renders the host significantly more susceptible tobleeding.

As a general proposition, the initial pharmaceutically effective amountof the Formula 1 compound administered parenterally per dose will be inthe range of about 0.01-100 mg/kg, namely about 0.1 to 20 mg/kg ofpatient body weight per day, with the typical initial range of compoundused being 0.3 to 15 mg/kg/day.

Acceptable diluents, carriers, excipients and stabilizers are nontoxicto recipients at the dosages and concentrations employed, and includebuffers such as phosphate, citrate and other organic acids; antioxidantsincluding ascorbic acid and methionine; preservatives (such asoctadecyldimethylbenzyl ammonium chloride; hexamethonium chloride;benzalkonium chloride, benzethonium chloride; phenol, butyl or benzylalcohol; alkyl parabens such as methyl or propyl paraben; catechol;resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecularweight (less than about 10 residues) polypeptides; proteins, such asserum albumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, histidine, arginine, or lysine; monosaccharides,disaccharides and other carbohydrates including glucose, mannose, ordextrins; chelating agents such as EDTA; sugars such as sucrose,mannitol, trehalose or sorbitol; salt-forming counter-ions such assodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionicsurfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG). Theactive pharmaceutical ingredients may also be entrapped in microcapsulesprepared, e.g., by coacervation techniques or by interfacialpolymerization, e.g., hydroxymethylcellulose or gelatin-microcapsulesand poly-(methylmethacylate) microcapsules, respectively, in colloidaldrug delivery systems (e.g., liposomes, albumin microspheres,microemulsions, nano-particles and nanocapsules) or in macroemulsions.Such techniques are disclosed in Remington's Pharmaceutical Sciences16^(th) edition, Osol, A. Ed. (1980).

Sustained-release preparations of Formula I compounds may be prepared.Suitable examples of sustained-release preparations includesemipermeable matrices of solid hydrophobic polymers containing acompound of Formula I, which matrices are in the form of shapedarticles, e.g., films, or microcapsules. Examples of sustained-releasematrices include polyesters, hydrogels (e.g.,poly(2-hydroxyethyl-methacrylate), or poly(vinyl alcohol)), polylactides(U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid andgamma-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate,degradable lactic acid-glycolic acid copolymers such as the LUPRONDEPOT™ (injectable microspheres composed of lactic acid-glycolic acidcopolymer and leuprolide acetate) and poly-D-(−)-3-hydroxybutyric acid.

The formulations include those suitable for the administration routesdetailed herein. The formulations may conveniently be presented in unitdosage form and may be prepared by any of the methods well known in theart of pharmacy. Techniques and formulations generally are found inRemington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa.).Such methods include the step of bringing into association the activeingredient with the carrier which constitutes one or more accessoryingredients. In general the formulations are prepared by uniformly andintimately bringing into association the active ingredient with liquidcarriers or finely divided solid carriers or both, and then, ifnecessary, shaping the product.

Formulations of a compound of Formula I suitable for oral administrationmay be prepared as discrete units such as pills, capsules, cachets ortablets each containing a predetermined amount of a compound of FormulaI.

Compressed tablets may be prepared by compressing in a suitable machinethe active ingredient in a free-flowing form such as a powder orgranules, optionally mixed with a binder, lubricant, inert diluent,preservative, surface active or dispersing agent. Molded tablets may bemade by molding in a suitable machine a mixture of the powdered activeingredient moistened with an inert liquid diluent. The tablets mayoptionally be coated or scored and optionally are formulated so as toprovide slow or controlled release of the active ingredient therefrom.

Tablets, troches, lozenges, aqueous or oil suspensions, dispersiblepowders or granules, emulsions, hard or soft capsules, e.g., gelatincapsules, syrups or elixirs may be prepared for oral use. Formulationsof compounds of Formula I intended for oral use may be preparedaccording to any method known to the art for the manufacture ofpharmaceutical compositions and such compositions may contain one ormore agents including sweetening agents, flavoring agents, coloringagents and preserving agents, in order to provide a palatablepreparation. Tablets containing the active ingredient in admixture withnon-toxic pharmaceutically acceptable excipient which are suitable formanufacture of tablets are acceptable. These excipients may be, e.g.,inert diluents, such as calcium or sodium carbonate, lactose, calcium orsodium phosphate; granulating and disintegrating agents, such as maizestarch, or alginic acid; binding agents, such as starch, gelatin oracacia; and lubricating agents, such as magnesium stearate, stearic acidor talc. Tablets may be uncoated or may be coated by known techniquesincluding microencapsulation to delay disintegration and adsorption inthe gastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate alone or with a wax may be employed.

For treatment of the eye or other external tissues, e.g., mouth andskin, the formulations may be applied as a topical ointment or creamcontaining the active ingredient(s) in an amount of, e.g., 0.075 to 20%w/w. When formulated in an ointment, the active ingredients may beemployed with either a paraffinic or a water-miscible ointment base.Alternatively, the active ingredients may be formulated in a cream withan oil-in-water cream base.

If desired, the aqueous phase of the cream base may include a polyhydricalcohol, i.e., an alcohol having two or more hydroxyl groups such aspropylene glycol, butane 1,3-diol, mannitol, sorbitol, glycerol andpolyethylene glycol (including PEG 400) and mixtures thereof. Thetopical formulations may desirably include a compound which enhancesabsorption or penetration of the active ingredient through the skin orother affected areas. Examples of such dermal penetration enhancersinclude dimethyl sulfoxide and related analogs.

The oily phase of the emulsions of this invention may be constitutedfrom known ingredients in a known manner. While the phase may comprisemerely an emulsifier, it desirably comprises a mixture of at least oneemulsifier with a fat or an oil or with both a fat and an oil.Preferably, a hydrophilic emulsifier is included together with alipophilic emulsifier which acts as a stabilizer. Together, theemulsifier(s) with or without stabilizer(s) make up the so-calledemulsifying wax, and the wax together with an oil and fat make up theso-called emulsifying ointment base which forms the oily dispersed phaseof the cream formulations. Emulsifiers and emulsion stabilizers suitablefor use in the formulation of the invention include Tween® 60, Span) 80,cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glycerylmono-stearate and sodium lauryl sulfate.

Aqueous suspensions of Formula I compounds contain the active materialsin admixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients include a suspending agent, such as sodiumcarboxymethylcellulose, croscarmellose, povidone, methylcellulose,hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone,gum tragacanth and gum acacia, and dispersing or wetting agents such asa naturally occurring phosphatide (e.g., lecithin), a condensationproduct of an alkylene oxide with a fatty acid (e.g., polyoxyethylenestearate), a condensation product of ethylene oxide with a long chainaliphatic alcohol (e.g., heptadecaethyleneoxycetanol), a condensationproduct of ethylene oxide with a partial ester derived from a fatty acidand a hexitol anhydride (e.g., polyoxyethylene sorbitan monooleate). Theaqueous suspension may also contain one or more preservatives such asethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one ormore flavoring agents and one or more sweetening agents, such as sucroseor saccharin.

The pharmaceutical compositions of compounds of Formula I may be in theform of a sterile injectable preparation, such as a sterile injectableaqueous or oleaginous suspension. This suspension may be formulatedaccording to the known art using those suitable dispersing or wettingagents and suspending agents which have been mentioned above. Thesterile injectable preparation may also be a sterile injectable solutionor suspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butanediol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils may conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid may likewise be used in the preparation ofinjectables.

The amount of active ingredient that may be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans maycontain approximately 1 to 1000 mg of active material compounded with anappropriate and convenient amount of carrier material which may varyfrom about 5 to about 95% of the total compositions (weight:weight). Thepharmaceutical composition can be prepared to provide easily measurableamounts for administration. For example, an aqueous solution intendedfor intravenous infusion may contain from about 3 to 500 μg of theactive ingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/hr can occur.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents.

Formulations suitable for topical administration to the eye also includeeye drops wherein the active ingredient is dissolved or suspended in asuitable carrier, especially an aqueous solvent for the activeingredient. The active ingredient is preferably present in suchformulations in a concentration of about 0.5 to 20% w/w, about 0.5 to10% w/w, or about 1.5% w/w.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouthwashes comprising the active ingredient in asuitable liquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising e.g. cocoa butter or a salicylate.

Formulations suitable for intrapulmonary or nasal administration have aparticle size e.g. in the range of 0.1 to 500 microns (includingparticle sizes in a range between 0.1 and 500 microns in incrementsmicrons such as 0.5, 1, 30 microns, 35 microns, etc.), which isadministered by rapid inhalation through the nasal passage or byinhalation through the mouth so as to reach the alveolar sacs. Suitableformulations include aqueous or oily solutions of the active ingredient.Formulations suitable for aerosol or dry powder administration may beprepared according to conventional methods and may be delivered withother therapeutic agents such as compounds heretofore used in thetreatment or prophylaxis disorders as described below.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

The formulations may be packaged in unit-dose or multi-dose containers,e.g. sealed ampoules and vials, and may be stored in a freeze-dried(lyophilized) condition requiring only the addition of the sterileliquid carrier, e.g. water, for injection immediately prior to use.Extemporaneous injection solutions and suspensions are prepared fromsterile powders, granules and tablets of the kind previously described.Preferred unit dosage formulations are those containing a daily dose orunit daily sub-dose, as herein above recited, or an appropriate fractionthereof, of the active ingredient.

The invention further provides veterinary compositions comprising atleast one active ingredient as above defined together with a veterinarycarrier therefore. Veterinary carriers are materials useful for thepurpose of administering the composition and may be solid, liquid orgaseous materials which are otherwise inert or acceptable in theveterinary art and are compatible with the active ingredient. Theseveterinary compositions may be administered parenterally, orally or byany other desired route.

Combination Therapy

The compounds of Formula I may be employed alone or in combination withother therapeutic agents for the treatment of a disease or disorderdescribed herein, such as a hyperproliferative disorder (e.g., cancer).In certain embodiments, a compound of Formula I is combined in apharmaceutical combination formulation, or dosing regimen as combinationtherapy, with a second compound that has anti-hyperproliferativeproperties or that is useful for treating a hyperproliferative disorder(e.g., cancer). The second compound of the pharmaceutical combinationformulation or dosing regimen preferably has complementary activities tothe compound of Formula I such that they do not adversely affect eachother. Such compounds are suitably present in combination in amountsthat are effective for the purpose intended. In one embodiment, acomposition of this invention comprises a compound of Formula I, incombination with a chemotherapeutic agent such as described herein.

The combination therapy may be administered as a simultaneous orsequential regimen. When administered sequentially, the combination maybe administered in two or more administrations. The combinedadministration includes coadministration, using separate formulations ora single pharmaceutical formulation, and consecutive administration ineither order, wherein preferably there is a time period while both (orall) active agents simultaneously exert their biological activities.

Suitable dosages for any of the above coadministered agents are thosepresently used and may be lowered due to the combined action (synergy)of the newly identified agent and other chemotherapeutic agents ortreatments.

The combination therapy may provide “synergy” and prove “synergistic”,i.e., the effect achieved when the active ingredients used together isgreater than the sum of the effects that results from using thecompounds separately. A synergistic effect may be attained when theactive ingredients are: (1) co-formulated and administered or deliveredsimultaneously in a combined, unit dosage formulation; (2) delivered byalternation or in parallel as separate formulations; or (3) by someother regimen. When delivered in alternation therapy, a synergisticeffect may be attained when the compounds are administered or deliveredsequentially, e.g., by different injections in separate syringes,separate pills or capsules, or separate infusions. In general, duringalternation therapy, an effective dosage of each active ingredient isadministered sequentially, i.e., serially, whereas in combinationtherapy, effective dosages of two or more active ingredients areadministered together.

In a particular embodiment of anti-cancer therapy, a compound of FormulaI, or a stereoisomer, geometric isomer, tautomer, solvate, metabolite,or pharmaceutically acceptable salt or prodrug thereof, may be combinedwith other chemotherapeutic, hormonal or antibody agents such as thosedescribed herein, as well as combined with surgical therapy andradiotherapy. Combination therapies according to the present inventionthus comprise it the administration of at least one compound of FormulaI, or a stereoisomer, geometric isomer, tautomer, solvate, metabolite,or pharmaceutically acceptable salt or prodrug thereof, and the use ofat least one other cancer treatment method. The amounts of thecompound(s) of Formula I and the other pharmaceutically activechemotherapeutic agent(s) and the relative timings of administrationwill be selected in order to achieve the desired combined therapeuticeffect.

Metabolites of Formula I Compounds

Also falling within the scope of this invention are the in vivometabolic products of Formula I described herein. Such products mayresult e.g. from the oxidation, reduction, hydrolysis, amidation,deamidation, esterification, deesterification, enzymatic cleavage, andthe like, of the administered compound. Accordingly, the inventionincludes metabolites of compounds of Formula I, including compoundsproduced by a process comprising contacting a compound of this inventionwith a mammal for a period of time sufficient to yield a metabolicproduct thereof.

Metabolite products typically are identified by preparing aradiolabelled (e.g., ¹⁴C or ³H) isotope of a compound of the invention,administering it parenterally in a detectable dose (e.g., greater thanabout 0.5 mg/kg) to an animal such as rat, mouse, guinea pig, monkey, orto man, allowing sufficient time for metabolism to occur (typicallyabout 30 seconds to 30 hours) and isolating its conversion products fromthe urine, blood or other biological samples. These products are easilyisolated since they are labeled (others are isolated by the use ofantibodies capable of binding epitopes surviving in the metabolite). Themetabolite structures are determined in conventional fashion, e.g., byMS, LC/MS or NMR analysis. In general, analysis of metabolites is donein the same way as conventional drug metabolism studies well known tothose skilled in the art. The metabolite products, so long as they arenot otherwise found in vivo, may be useful in diagnostic assays fortherapeutic dosing of the compounds of the invention.

Articles of Manufacture

In another embodiment of the invention, an article of manufacture, or“kit”, containing materials useful for the treatment of the diseases anddisorders described above is provided. The kit comprises a containercomprising a compound of Formula I. The kit may further comprise a labelor package insert, on or associated with the container. The term“package insert” is used to refer to instructions customarily includedin commercial packages of therapeutic products, that contain informationabout the indications, usage, dosage, administration, contraindicationsand/or warnings concerning the use of such therapeutic products.

One embodiment of the invention comprises a kit for treating aPI3K-mediated condition, comprising a compound of this invention, andinstructions for use.

Suitable containers include, e.g., bottles, vials, syringes, blisterpack, etc. The container may be formed from a variety of materials suchas glass or plastic. The container may hold a compound of Formula I or aformulation thereof which is effective for treating the condition andmay have a sterile access port (e.g., the container may be anintravenous solution bag or a vial having a stopper pierceable by ahypodermic injection needle). At least one active agent in thecomposition is a compound of Formula I. The label or package insertindicates that the composition is used for treating the condition ofchoice, such as cancer. In addition, the label or package insert mayindicate that the patient to be treated is one having a disorder such asa hyperproliferative disorder, neurodegeneration, cardiac hypertrophy,pain, migraine or a neurotraumatic disease or event. In one embodiment,the label or package inserts indicates that the composition comprising acompound of Formula I can be used to treat a disorder resulting fromabnormal cell growth. The label or package insert may also indicate thatthe composition can be used to treat other disorders. Alternatively, oradditionally, the article of manufacture may further comprise a secondcontainer comprising a pharmaceutically acceptable buffer, such asbacteriostatic water for injection (BWFI), phosphate-buffered saline,Ringer's solution and dextrose solution. It may further include othermaterials desirable from a commercial and user standpoint, includingother buffers, diluents, filters, needles, and syringes.

The kit may further comprise directions for the administration of thecompound of Formula I and, if present, the second pharmaceuticalformulation. For example, if the kit comprises a first compositioncomprising a compound of Formula I, and a second pharmaceuticalformulation, the kit may further comprise directions for thesimultaneous, sequential or separate administration of the first andsecond pharmaceutical compositions to a patient in need thereof.

In another embodiment, the kits are suitable for the delivery of solidoral forms of a compound of Formula I, such as tablets or capsules. Sucha kit preferably includes a number of unit dosages. Such kits caninclude a card having the dosages oriented in the order of theirintended use. An example of such a kit is a “blister pack”. Blisterpacks are well known in the packaging industry and are widely used forpackaging pharmaceutical unit dosage forms. If desired, a memory aid canbe provided, e.g. in the form of numbers, letters, or other markings orwith a calendar insert, designating the days in the treatment schedulein which the dosages can be administered.

According to one embodiment, a kit may comprise (a) a first containerwith a compound of Formula I contained therein; and optionally (b) asecond container with a second pharmaceutical formulation containedtherein, wherein the second pharmaceutical formulation comprises asecond compound with anti-hyperproliferative activity. Alternatively, oradditionally, the kit may further comprise a third container comprisinga pharmaceutically-acceptable buffer, such as bacteriostatic water forinjection (BWFI), phosphate-buffered saline, Ringer's solution anddextrose solution. It may further include other materials desirable froma commercial and user standpoint, including other buffers, diluents,filters, needles, and syringes.

In certain other embodiments wherein the kit comprises a composition ofFormula I and a second therapeutic agent, the kit may comprise acontainer for containing the separate compositions such as a dividedbottle or a divided foil packet, however, the separate compositions mayalso be contained within a single, undivided container. Typically, thekit comprises directions for the administration of the separatecomponents. The kit form is particularly advantageous when the separatecomponents are preferably administered in different dosage forms (e.g.,oral and parenteral), are administered at different dosage intervals, orwhen titration of the individual components of the combination isdesired by the prescribing physician.

Preparation of Formula I Compounds

Tricyclic compounds of Formula I may be synthesized by synthetic routesthat include processes analogous to those well-known in the chemicalarts, particularly in light of the description contained herein. Thestarting materials are generally available from commercial sources suchas Aldrich Chemicals (Milwaukee, Wis.) or are readily prepared usingmethods well known to those skilled in the art (e.g., prepared bymethods generally described in Louis F. Fieser and Mary Fieser, Reagentsor Organic Synthesis, v. 1-23, Wiley, N.Y. (1967-2006 ed.), orBeilsteins Handbuch der organischen Chemie, 4, Aufl. ed.Springer-Verlag, Berlin, including supplements (also available via theBeilstein online database).

In certain embodiments, compounds of Formula I may be readily preparedusing well-known procedures to prepare purine compounds (Hammarstrom etal (2007) Tetrahedron Lett. 48(16):2823-2827; Cerna et al (2006) OrganicLetters 8(23):5389-5392; Chang et al (2006) J. Med. Chem.49(10):2861-2867; Yang et al (2005) J. Comb. Chem. 7:474-482; Liu et al(2005) J. Comb. Chem. 7:627-636; Hocek et al (2004) Synthesis17:2869-2876; Hammarstrom et al (2003) Tetrahedron Lett. 44:8361-8363;Hammarstrom et al (2002) Tetrahedron Lett. 43:8071-8073; Booth et al(1987) J. Chem. Soc, Perkin Trans. 1: Organic and Bio-Organic Chem.7:1521-1526; Booth et al (1981) J. Chem. Soc., Chemical Communications15:788-789; Yoneda et al (1976) J. Chem. Soc., Perkin Trans. 1: Organicand Bio-Organic Chem. 14:1547-1550; Taylor et al (1971) J. Org. Chem.36(21):3211-3217; Lister, J. H.; Fenn, M. D. The Purines, Supplementary1, John Wiley & Sons, 1996, Volume 54; The Chemistry of HeterocyclicCompounds, Editors Weissberger, A.; Taylor E. C., Wiley Interscience,1971, Volume 24; Legraverend, M.; Grierson, D. S. (2006) Bioorg. Med.Chem. 14:3987-4006; Hocek, M. (2003) Eur. J. Org. Chem. 245-254; U.S.Pat. Nos. 7,122,665; 6,743,919; 5,332,744; 4,728,644; 3,016,378; US2008/0058297; US 2003/0139427; WO 2008/043031); and other heterocycles,which are described in: Comprehensive Heterocyclic Chemistry II, EditorsKatritzky and Rees, Elsevier, 1997, e.g. Volume 3; Liebigs Annalen derChemie, (9):1910-16, (1985); Helvetica Chimica Acta, 41:1052-60, (1958);Arzneimittel-Forschung, 40(12):1328-31, (1990).

Compounds of Formula I may be prepared singly or as compound librariescomprising at least 2, e.g. 5 to 1,000 compounds, or 10 to 100compounds. Libraries of compounds of Formula I may be prepared by acombinatorial ‘split and mix’ approach or by multiple parallel synthesesusing either solution phase or solid phase chemistry, by proceduresknown to those skilled in the art. Thus according to a further aspect ofthe invention there is provided a compound library comprising at least 2compounds, or pharmaceutically acceptable salts thereof.

For illustrative purposes, the General Procedures show general methodswhich may be applied for preparation of Formula I compounds, as well askey intermediates. The FIGURES and Examples sections contain moredetailed description of individual reaction steps. Those skilled in theart will appreciate that other synthetic routes may be used tosynthesize the inventive compounds. Although certain starting materialsand routes are depicted in the Schemes, General Procedures and Examples,other similar starting materials and routes can be substituted toprovide a variety of derivatives and/or reaction conditions. Inaddition, many of the compounds prepared by the methods described belowcan be further modified in light of this disclosure using conventionalchemistry well known to those skilled in the art.

In preparing compounds of Formula I, protection of remote functionality(e.g., primary or secondary amine) of intermediates may be necessary.The need for such protection will vary depending on the nature of theremote functionality and the conditions of the preparation methods.Suitable amino-protecting groups include acetyl, trifluoroacetyl,t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBz) and9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection isreadily determined by one skilled in the art. For a general descriptionof protecting groups and their use, see T. W. Greene, Protective Groupsin Organic Synthesis, John Wiley & Sons, New York, Third Ed., 1999.

Methods of Separation

In the methods of preparing the compounds of this invention, it may beadvantageous to separate reaction products from one another and/or fromstarting materials. The desired products of each step or series of stepsis separated and/or purified (hereinafter separated) to the desireddegree of homogeneity by the techniques common in the art. Typicallysuch separations involve multiphase extraction, crystallization from asolvent or solvent mixture, distillation, sublimation, orchromatography. Chromatography can involve any number of methodsincluding, e.g.: reverse-phase and normal phase; size exclusion; ionexchange; high, medium and low pressure liquid chromatography methodsand apparatus; small scale analytical; simulated moving bed (SMB) andpreparative thin or thick layer chromatography, as well as techniques ofsmall scale thin layer and flash chromatography.

Another class of separation methods involves treatment of a mixture witha reagent selected to bind to or render otherwise separable a desiredproduct, unreacted starting material, reaction by product, or the like.Such reagents include adsorbents or absorbents such as activated carbon,molecular sieves, ion exchange media, or the like. Alternatively, thereagents can be acids in the case of a basic material, bases in the caseof an acidic material, binding reagents such as antibodies, bindingproteins, selective chelators such as crown ethers, liquid/liquid ionextraction reagents (LIX), or the like.

Selection of appropriate methods of separation depends on the nature ofthe materials involved. For example, boiling point and molecular weightin distillation and sublimation, presence or absence of polar functionalgroups in chromatography, stability of materials in acidic and basicmedia in multiphase extraction, and the like. One skilled in the artwill apply techniques most likely to achieve the desired separation.

Diastereomeric mixtures can be separated into their individualdiastereomers on the basis of their physical chemical differences bymethods well known to those skilled in the art, such as bychromatography and/or fractional crystallization. Enantiomers can beseparated by converting the enantiomeric mixture into a diastereomericmixture by reaction with an appropriate optically active compound (e.g.,chiral auxiliary such as a chiral alcohol or Mosher's acid chloride),separating the diastereomers and converting (e.g., hydrolyzing) theindividual diastereoisomers to the corresponding pure enantiomers. Also,some of the compounds of the present invention may be atropisomers(e.g., substituted biaryls) and are considered as part of thisinvention. Enantiomers can also be separated by use of a chiral HPLCcolumn.

A single stereoisomer, e.g., an enantiomer, substantially free of itsstereoisomer may be obtained by resolution of the racemic mixture usinga method such as formation of diastereomers using optically activeresolving agents (Eliel, E. and Wilen, S. “Stereochemistry of OrganicCompounds,” John Wiley & Sons, Inc., New York, 1994; Lochmuller, C. H.,(1975) J. Chromatogr., 113(3):283-302). Racemic mixtures of chiralcompounds of the invention can be separated and isolated by any suitablemethod, including: (1) formation of ionic, diastereomeric salts withchiral compounds and separation by fractional crystallization or othermethods, (2) formation of diastereomeric compounds with chiralderivatizing reagents, separation of the diastereomers, and conversionto the pure stereoisomers, and (3) separation of the substantially pureor enriched stereoisomers directly under chiral conditions. See: “DrugStereochemistry, Analytical Methods and Pharmacology,” Irving W. Wainer,Ed., Marcel Dekker, Inc., New York (1993).

Under method (1), diastereomeric salts can be formed by reaction ofenantiomerically pure chiral bases such as brucine, quinine, ephedrine,strychnine, α-methyl-β-phenylethylamine (amphetamine), and the like withasymmetric compounds bearing acidic functionality, such as carboxylicacid and sulfonic acid. The diastereomeric salts may be induced toseparate by fractional crystallization or ionic chromatography. Forseparation of the optical isomers of amino compounds, addition of chiralcarboxylic or sulfonic acids, such as camphorsulfonic acid, tartaricacid, mandelic acid, or lactic acid can result in formation of thediastereomeric salts.

Alternatively, by method (2), the substrate to be resolved is reactedwith one enantiomer of a chiral compound to form a diastereomeric pair(E. and Wilen, S. “Stereochemistry of Organic Compounds”, John Wiley &Sons, Inc., 1994, p. 322). Diastereomeric compounds can be formed byreacting asymmetric compounds with enantiomerically pure chiralderivatizing reagents, such as menthyl derivatives, followed byseparation of the diastereomers and hydrolysis to yield the pure orenriched enantiomer. A method of determining optical purity involvesmaking chiral esters of the racemic mixture, such as a menthyl ester,for example with (−) menthyl chloroformate, in the presence of base, orMosher ester, α-methoxy-α-(trifluoromethyl)phenyl acetate (Jacob II. J.Org. Chem. (1982) 47:4165), and analyzing the ¹H NMR spectrum for thepresence of the two atropisomeric enantiomers or diastereomers. Stablediastereomers of atropisomeric compounds can be separated and isolatedby normal- and reverse-phase chromatography following methods forseparation of atropisomeric naphthyl-isoquinolines (WO 1996/015111). Bymethod (3), a racemic mixture of two enantiomers can be separated bychromatography using a chiral stationary phase (“Chiral LiquidChromatography” (1989) W. J. Lough, Ed., Chapman and Hall, New York;Okamoto, J. Chromatogr., (1990) 513:375-378). Enriched or purifiedenantiomers can be distinguished by methods used to distinguish otherchiral molecules with asymmetric carbon atoms, such as optical rotationand circular dichroism.

General Preparative Procedures General Procedure A Suzuki Coupling:

The Suzuki-type coupling reaction is useful to attach a monocyclicheteroaryl, a fused bicyclic heterocycle, a fused bicyclic heteroaryl,or a phenyl at the 2-position of the pyrimidine ring of a2-chloro-purine 21. For example, 21 may be combined with 1.5 equivalentsof 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)1H-indazole 24, anddissolved in about 3 equivalents of sodium carbonate as about a 1 molarsolution in water and about an equal volume of acetonitrile. A catalyticamount, or more, of a low valent palladium reagent, such asbis(triphenylphosphine)palladium(II) dichloride, is added. A variety ofboronic acids or boronic esters can be used in place of the indazoleboronic ester indicated. Also alternatively, the nitrogen of theindazole may be protected, for example, N-THP protected compound 41. Insome cases potassium acetate was used in place of sodium carbonate toadjust the pH of the aqueous layer. The Suzuki palladium couplingreaction may be optimized and/or accelerated under microwave conditions.The reaction may be heated at about 100-150° C. under pressure in amicrowave reactor such as the Biotage Optimizer (Biotage, Inc.) forabout 10 to 30 minutes. The contents are cooled, concentrated, andextracted with ethyl acetate, or another organic solvent. Afterevaporation of the organic layer the Suzuki coupling products,6,8,9-substituted 2-(1H-indazol-4-yl)-purine 22, or 6,8,9-substituted2-(5-pyrimidin-2-amine)-purine 23, may be purified on silica or byreverse phase HPLC. Substituents R^(2′), R^(3′) may be R², R³ asdefined, or protected forms or precursors thereof.

A variety of palladium catalysts can be used during the Suzuki couplingstep to form compounds, including exemplary embodiments 22 and 23.Suzuki coupling is a palladium mediated cross coupling reaction of anarylhalide, such as 21, with a boronic acid or ester such as4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole24 or5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine 25. Lowvalent, Pd(II) and Pd(0) catalysts may be used in the Suzuki couplingreaction, including PdCl2(PPh₃)₂, Pd(t-Bu)₃, PdCl₂ dppf CH₂Cl₂,Pd(PPh₃)₄, Pd(Oac)/PPh₃, Cl₂Pd[(Pet₃)]₂, Pd(DIPHOS)₂, Cl₂Pd(Bipy),[PdCl(Ph₂PCH₂PPh₂)]₂, Cl₂Pd[P(o-tol)₃]₂, Pd₂(dba)₃/P(o-tol)₂,Pd₂(dba)/P(furyl)₃, Cl₂Pd[P(furyl)₃]₂, Cl₂Pd(PmePh₂)₂,Cl₂Pd[P(4-F-Ph)₃]₂, Cl₂Pd[P(C₆F₆)₃]₂, Cl₂Pd[P(2-COOH-Ph)(Ph)₂]₂,Cl₂Pd[P(4-COOH-Ph)(Ph)₂]₂, and encapsulated catalysts Pd EnCat™ 30, PdEnCat™ TPP30, and Pd(II)EnCat™ BINAP30 (US 2004/0254066). One suchSuzuki palladium catalyst is[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complexwith dichloromethane, represented as Pd (dppf)Cl₂.

General Procedure B C-6 Nitrogen Substitution

To a 2,6-dichloro purine intermediate 27 in a solvent such as ethanol isadded a morpholino amine (mor, 1.1 equiv.) and a non-nucleophilic basesuch as triethylamine (Net₃, 1.5 equiv.). Alternatively, acetonitrilemay be used as the solvent and potassium carbonate may be used as thebase. The reaction mixture is stirred at room temperature for about 1hour or overnight, volatiles removed in vacuo and residue partitionedbetween DCM and brine. If the mixture is insoluble it may be sonicatedand the solid product was collected by filtration. Drying with magnesiumsulfate and evaporation of the solvent gives N′-(2-chloropurin-6-yl)-amine substituted intermediate 28, often as a crystallinesolid, or by trituration. Substituents R^(2′) and R^(3′) may be R² andR³ as defined, or protected forms or precursors thereof.

General Procedure C N-9 Nitrogen Alkylation

9-H Purine intermediate 29 is brought up into DMF and 2 equiv of cesiumcarbonate is added to the reaction mixture. The reaction is heated to50° C. whereupon 3 equivalents of an alkyl halide R^(2′)—X are added tothe reaction mixture. The reaction is monitored by TLC or LC/MS andstirred until completion, typically several hours. The reaction mixtureis extracted with EtOAc and water, and the organic layer is dried,filtered and concentrated to get crude 9-alkylated purine 30 which isused directly in the next reaction or purified by reverse phase HPLC.Substituents R², R³ and R⁴ may be R², R³ and R⁴ as defined, or protectedforms or precursors thereof.

General Procedure D THP Deprotection

Generally, N-9-tetrahydropyranyl substituted 31 may be treated withcatalytic amounts of para-toluenesulfonic acid (PTSA) in a solution ofmethanol and heated to about 50° C. until the tetrahydropyran (THP)group is removed to afford compound 32. The reaction may be monitored byLC-MS or TLC. Substituents R^(3′) and R^(4′) may be R³ and R⁴ asdefined, or protected forms or precursors thereof.

General Procedure E Boc Deprotection

Generally, Boc-substituted 33 is treated with TFA or 4N HCl to removethe t-butoxycarbonyl group(s) and the reaction is monitored by LC-MS forcompletion. The crude product is then concentrated and purified byreverse phase HPLC to yield product 34 as a pure solid. SubstituentsR^(2′) and R^(3′) may be R² and R³ as defined, or protected forms orprecursors thereof.

General Procedure F Amide Coupling

A 2,6,8 substituted, 9-alkylcarboxyl purine 35, where n is 1 to 3, istreated with 1.5 eq HATU(2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate), an excess (such as 3 eq) of an alkylamine(HNR¹⁰R¹¹) and an excess (such as 3 eq) of cesium carbonate indimethylformamide (DMF). Alternatively, other coupling reagents may beused. The reaction is stirred until complete and extracted inethylacetate with saturated bicarbonate solution. The organic layer isdried, filtered and concentrated to yield the acylated, crudeintermediate, which is purified via reverse phase HPLC to yield product36. Substituents R^(3′) and R^(4′) may be R³ and R⁴ as defined, orprotected forms or precursors thereof.

General Procedure G Pyrimidooxazine (Formula In) synthesis

The pyrimidooxazines of formula (In) are prepared according to theprocedures described below in Scheme 1 above, or by methods known in theart. Protected trichloropyrimidines of formula (II) may be preparedusing methods described in the literature. The trichlorides may bereacted with a cyclic amine of formula (IIA) in the presence of a basesuch as triethylamine in a solvent such as ethanol at around ambienttemperature to give dichlorides of formula (III). The methoxy pyrimidinemay be deprotected using a reagent such as LiCl in a solvent such as DMFunder microwave irradiation for PG=Me or by treatment with an acid suchas TFA in a solvent such as DCM when PG=p-methoxybenzyl to give phenolsof formula (IV). Tricyclic pyrimidooxazines of formula (V) may then beformed from diols of formula (IV) with a azo compound such as DIAD inthe presence of a phosphine such as triphenyl phosphine in a solventsuch as 1,4-dioxane. Alternatively, formation of compounds (V) may beaccomplished by firstly transforming the hydroxyl group into a suitableleaving group followed by an intramolecular substitution reactionfacilitated by reaction with a reagent such as methanesulfonyl chloridein the presence of base such as triethylamine in a solvent such as THF.Compounds of formula (VI) may be prepared from compounds (V) by reactionwith a morpholine derivative (incorporating appropriate substituents R)in the presence of a base such as triethylamine in a solvent such asethanol at elevated temperature. For n=2, A=O, the addition ofmorpholine occurs in a regiospecific manner whereas for n=1, A=CH₂, thisreaction may also lead to the formation of the undesired regioisomer.Pyrimidooxazines of formula (1) may be formed by reaction of compoundsof formula (VI) with an aryl or heteroaryl metallated reactant such as aheteroaryl boronic acid, boronic ester or a stannane in the presence ofa transition metal catalyst such as Pd(PPh₃)₂Cl₂ and a base such asaqueous sodium carbonate in a solvent such as acetonitrile undermicrowave irradiation at a temperatures of up to 150° C.

Alternatively, pyrimidooxazines of formula (In) may be preparedaccording to Scheme 2 below. Protected trichloro pyrimidines of formula(II) may be obtained from compounds of formula (VII) (prepared accordingto methods described in the art) by reaction with an alcohol such asp-methoxybenzyl alcohol in the presence of a azo compound such as DIADin the presence of a phosphine such as triphenyl phosphine in a solventsuch as 1,4-dioxane. Alternatively, formation of compounds (II) may beaccomplished by treatment of compound (VII) with a silyl halide such astert-butylchlorodiphenylsilane in the presence of a base such astriethylamine with as additive such as N,N-dimethylaminopyridine in asolvent as DMF. The trichloropyrimidines (II) may be reacted with acyclic amine of formula (IIA) in the presence of a base such astriethylamine in a solvent such as ethanol at ambient temperature togive dichlorides of formula (VII). Compounds of formula (IX) may beprepared by reaction of compounds of formula (VIII) with an aryl orheteroaryl metallated reactant such as a heteroaryl boronic acid,boronic ester or a stannane in the presence of a transition metalcatalyst such as Pd(PPh₃)₂Cl₂ and a base such as aqueous sodiumcarbonate in a solvent such as acetonitrile under microwave irradiationat a temperatures of up to 150° C. For bulky protection groups, e.g.PG=p-methoxybenzene or PG=tert-butyldiphenylsilane, this substitutionmay occur with regiospecific control giving 2-aryl or 2-heteroarylcompounds of formula (IX). Phenols of formula (X) may be formed bydeprotection of compounds (IX) using an acid such as TFA in a solventsuch as DCM at ambient temperature. Compounds of formula (X) may beconverted to compounds of formula (XI) using the methods described forthe conversion of compounds of formula (IV) to compounds of formula (V)in Scheme 1. Finally, pyrimidooxazines of formula (In) may formed byreaction of compounds of formula (XI) by reaction with a morpholine(incorporating appropriate substituents R) in the presence of a basesuch as triethylamine in a solvent such as ethanol at temperatures up toreflux.

EXAMPLES

The chemical reactions described in the Examples may be readily adaptedto prepare a number of other PI3K inhibitors of the invention, andalternative methods for preparing the compounds of this invention aredeemed to be within the scope of this invention. For example, thesynthesis of non-exemplified compounds according to the invention may besuccessfully performed by modifications apparent to those skilled in theart, e.g., by appropriately protecting reactive functional groups, byutilizing other suitable reagents known in the art other than thosedescribed, and/or by making routine modifications of reactionconditions. Alternatively, other reactions disclosed herein or known inthe art will be recognized as having applicability for preparing othercompounds of the invention.

In the Examples described below, unless otherwise indicated alltemperatures are set forth in degrees Celsius. Reagents were purchasedfrom commercial suppliers such as Sigma Aldrich Chemical Company,Lancaster, TCI or Maybridge, and were used without further purificationunless otherwise indicated. The reactions set forth below were donegenerally under a positive pressure of nitrogen or argon or with adrying tube (unless otherwise stated) in anhydrous solvents, and thereaction flasks were typically fitted with rubber septa for theintroduction of substrates and reagents via syringe. Glassware was ovendried and/or heat dried. Column chromatography was conducted on aBiotage system (Manufacturer: Dyax Corporation) having a silica gelcolumn or on a silica SEP PAK® cartridge (Waters). ¹H NMR spectra wereobtained at 400 MHz in deuterated CDCl₃, d₆-DMSO, CH₃OD or d₆-acetonesolutions (reported in ppm), using chloroform as the reference standard(7.25 ppm). When peak multiplicities are reported, the followingabbreviations are used: s (singlet), d (doublet), t (triplet), m(multiplet), br (broadened), dd (doublet of doublets), dt (doublet oftriplets). Coupling constants, when given, are reported in hertz (Hz).

HPLC was conducted by the following exemplary methods:

(A) LCMS short method—10 min run

HPLC-Agilent 1200 Mobile phase A Water with 0.05% TFA Mobile phase BAcetonitrile with 0.05% TFA Agilent ZORBAX SD-C18, 1.8 μm, Column 2.1 ×30 mm Column temperature 40 degree C. LC gradient 3-95% B in 8.5 min,95% in 2.5 min LC Flowrate 400 μL/min UV wavelength 220 nm and 254 nmMass Spec - Agilent quadrupole 6140 Ionization ESI positive Scan range110-800 amu(B) Waters Acquity/LCT long method—20 min run

Waters Acquity UPLC Mobile phase A Waters with 0.05% TFA Mobile phase BAcetonitrile with 0.05% TFA Acquity UPLC BEH C18, 1.7 μm, Column 2.1 ×50 mm Column temperature 40 degree C. LC gradient 2-98% B in 17.0 min,98% in 1.5 min LC Flowrate 600 μL/min UV wavelength 254 nm Mass Spec -Waters LCT Premier XE Ionization ESI positive Scan range 100-800 amu(C) LCMS 2.5 min Chiral method

Mobile Phase A: CO2

Mobile Phase B: methanolIsocratic conditions: 25% BFlow rate: 5 mL/minOutlet pressure: 120 Bar

Temperature: 40° C. Column: ChiralCel OJ (4.6×50 mm, 3 μm) Uv: 230 nmSystem: Berger Analytical SFC/MS

Chiral purification:

Conditions A: Mobile Phase A: CO2

Mobile Phase B: methanolIsocratic conditions: 25% BFlow rate: 60 mL/minOutlet pressure: 100 BarTemperature: 40 degrees C.

Column: ChiralCel OJ (21.2×250 mm, 5 μm) Uv: 230 nm System: Berger MGIIExample 1 2,6-dichloro-9-methyl-9H-purine 4

The cyano group of 5-amino-1-methyl-1H-imidazole-4-carbonitrile 1 ishydrolyzed to the amide in sulfuric acid to give5-amino-1-methyl-1H-imidazole-4-carboxamide 2 which was cyclized withurea to 9-methyl-1H-purine-2,6(3H,9H)-dione 3. Chlorination of 3 yields2,6-dichloro-9-methyl-9H-purine 4.

Example 2 4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indazole24—route 1

To a solution of 3-bromo-2-methyl aniline (5.0 g, 26.9 mmol) inchloroform (50 mL) was added potassium acetate (1.05 eq., 28.2 mmol,2.77 g). Acetic anhydride (2.0 eq., 53.7 mmol, 5.07 mL) was added withconcurrent cooling in ice-water. The mixture was then stirred at roomtemperature for 10 minutes after which time a white gelatinous solidformed. 18-Crown-6 (0.2 eq., 5.37 mmol, 1.42 g) was added followed byiso-amyl nitrite (2.2 eq., 59.1 mmol, 7.94 mL) and the mixture washeated under reflux for 18 h. The reaction mixture was allowed to cool,and was partitioned between chloroform (3×100 mL) and saturated aqueoussodium hydrogen carbonate (100 mL). The combined organic extracts werewashed with brine (100 mL), separated and dried (MgSO₄). The crudeproduct was evaporated onto silica and purified by chromatographyeluting with 20% to 40% EtOAc-petrol to give1-(4-bromo-indazol-1-yl)-ethanone A (3.14 g, 49%) as an orange solid,and 4-bromo-1H-indazole B (2.13 g, 40%) as a pale orange solid. A: ¹HNMR (400 MHz, CDCl₃) 2.80 (3H, s), 7.41 (1H, t, J=7.8 Hz), 7.50 (1H, d,J=7.8 Hz), 8.15 (1H, s), 8.40 (1H, d, J=7.8 Hz). B: ¹H NMR (400 MHz,CDCl₃) 7.25 (1H, t, J=7.3 Hz), 7.33 (1H, d, J=7.3 Hz), 7.46 (1H, d,J=7.3 Hz), 8.11 (1H, s), 10.20 (1H, br s).

To a solution of the 1-(4-bromo-indazol-1-yl)-ethanone A (3.09 g, 12.9mmol) in MeOH (50 mL) was added 6N aqueous HCl (30 mL) and the mixturewas stirred at room temperature for 7 h. The MeOH was evaporated and themixture partitioned between EtOAc (2×50 mL) and water (50 mL). Thecombined organic layers were washed with brine (50 mL), separated anddried (MgSO₄). The solvent was removed by evaporation under reducedpressure to give 4-bromo-1H-indazole B (2.36 g, 93%).

To a solution of the 4-bromo-1H-indazole B (500 mg, 2.54 mmol) andbis(pinacolato)diboron (1.5 eq., 3.81 mmol) in DMSO (20 mL) was addedpotassium acetate (3.0 eq., 7.61 mmol, 747 mg; dried in drying pistol)and PdCl₂(dppf)₂ (3 mol %, 0.076 mmol, 62 mg). The mixture was degassedwith argon and heated at 80° C. for 40 h. The reaction mixture wasallowed to cool and partitioned between water (50 mL) and ether (3×50mL). The combined organic layers were washed with brine (50 mL),separated and dried (MgSO₄). The crude material was purified bychromatography eluting with 30% to 40% EtOAc-petrol to give aninseparable 3:1 mixture of the4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indazole 24 (369 mg,60%) and indazole (60 mg, 20%), isolated as a yellow gum whichsolidified upon standing to furnish as an off-white solid. ¹H NMR (400MHz, d₆-DMSO) 1.41 (12H, s), 7.40 (1H, dd, J=8.4 Hz, 6.9 Hz), 7.59 (1H,d, J=8.4 Hz), 7.67 (1H, d, J=6.9 Hz), 10.00 (1H, br s), 8.45 (1H, s),and indazole: 7.40 (1H, t), 7.18 (1H, t, J=7.9 Hz), 7.50 (1H, d, J=9.1Hz), 7.77 (1H, d, J=7.9 Hz), 8.09 (1H, s); impurity at 1.25.

Example 3 4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indazole24—route 2

To a solution of 2-methyl-3-nitroaniline (2.27 g, 14.91 mmol) in aceticacid (60 mL) was added a solution of sodium nitrite (1.13 g, 1.1 eq.) inwater (5 mL). After 2 h, the deep red solution was poured onto ice/waterand the resulting precipitate collected by filtration to yield4-nitro-1H-indazole C (1.98 g, 81%).

A mixture of 4-nitro-1H-indazole C (760 mg, 4.68 mmol), palladium oncharcoal (10%, cat.) and ethanol (30 m L) was stirred under a balloon ofhydrogen for 4 h. The reaction mixture was then filtered through celite,and the solvent removed in vacuo to yield 1H-indazol-4-ylamine D (631mg, 100%).

An aqueous solution of sodium nitrite (337 mg, 4.89 mmol) in water (2mL) was added dropwise to a suspension of 1H-indazol-4-ylamine D (631mg, 4.74 mmol) in 6M hydrochloric acid (7.2 mL) at below 0° C. Afterstirring for 30 minutes, sodium tetrafluoroborate (724 mg) was added tothe reaction mixture. A viscous solution resulted, which was filteredand washed briefly with water to yield 1H-indazole-4-diazoniumtetrafluoroborate salt E (218 mg, 20%) as a deep red solid.

Dry methanol (4 mL) was purged with argon for 5 minutes. To this wasadded 1H-indazole-4-diazonium tetrafluoroborate salt (218 mg, 0.94mmol), bis-pinacolato diboron (239 mg, 1.0 eq.) and[1,1′-bis(diphenylphosphino)ferrocene]palladium (II) chloride (20 mg).

The reaction mixture was stirred for 5 h and then filtered throughcelite. The residue was purified using flash chromatography to yield4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indazole 24 (117mg).

Example 41-(Tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole(Route A)

Step A: Preparation of 4-chloro-1H-indazole: To a 250 ml flask with stirbar was added 2-methyl-3-chloroaniline (8.4 ml, 9.95 g, 70.6 mmol),potassium acetate (8.3 g, 84.7 mmol) and chloroform (120 ml). Thismixture was cooled to 0° C. with stirring. To the cooled mixture wasadded acetic anhydride (20.0 ml, 212 mmol) drop wise over 2 minutes. Thereaction mixture was warmed to 25° C. and stirred for 1 hour. At thispoint, the reaction was heated to 60° C. Isoamyl nitrite (18.9 ml, 141mmol) was added and the reaction was stirred overnight at 60° C. Oncecomplete, water (75 ml) and THF (150 ml) were added and the reaction wascooled to 0° C. LiOH (20.7 g, 494 mmol) was added and the reaction wasstirred at 0° C. for 3 hours. Water (200 ml) was added and the productwas extracted with EtOAc (300 ml, 100 ml). The organic layers werecombined, dried with MgSO₄ and concentrated in vacuo to yield4-chloro-1H-indazole 11.07 g (100%) as an orange solid. ¹H NMR (400 MHz,CDCl₃) δ 8.18 (d, J=1 Hz, 1H), 7.33 (d, J=8 Hz 1H), 7.31 (t, J=7 Hz,1H), 7.17 (dd, J=7 Hz, 1 Hz, 1H). LCMS (ESI pos) m/c 153 (M+1).

Step B: Preparation of4-chloro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole: To a 1 L flask withmechanical stirrer was added 4-chloro-1H-indazole (75.0 g, 0.492 mol),pyridinium p-toluenesulfonate (1.24 g, 4.92 mmol), CH₂Cl₂ (500 ml) and3,4-dihydro-2H-pyran (98.6 ml, 1.08 mol). With stirring, this mixturewas heated to 45° C. for 16 hours. Analysis of reaction mixture showsproduction of both isomers of product. Cooled reaction to 25° C. andadded CH₂Cl₂ (200 ml). Washed the solution with water (300 mil) andsaturated NaHCO₃ (250 ml). Dried the organics with MgSO₄ andconcentrated to dryness. Purified the crude product by dissolving inEtOAc/hexanes (4:6, 1 L) and adding SiO₂ (1.2 L). The mixture wasfiltered and the cake was washed with EtOAc/Hexanes (4:6, 2 L). Theorganics were concentrated in vacuo to yield4-chloro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole 110.2 g (95%) as anorange solid. Isomer 1: ¹H NMR (400 MHz, CDCl₃) δ 8.10 (d, J=1 Hz, 1H),7.50 (dd, J=9 Hz, 1 Hz, 1H), 7.29 (dd, J=9 Hz, 8 Hz, 1H), 7.15 (dd, J=8Hz, 1 Hz 1H) 5.71 (dd, J=9 Hz, 3 Hz, 1H) 4.02 (m, 1H) 3.55 (m, 1H) 2.51(m, 1H) 2.02 (m, 2H) 1.55 (m, 3H). LCMS (ESI pos) m/c 237 (M+1); Isomer2: ¹H NMR (400 MHz, CDCl₃) δ 8.25 (d, J=1 Hz, 1H), 7.62 (dd, J=9 Hz, 1Hz 1H), 7.20 (dd, J=9 Hz, 8 Hz 1H), 7.06 (dd, J=8 Hz, 1 Hz 1H) 5.69 (dd,J=9 Hz, 3 Hz 1H) 4.15 (m, 1H) 3.80 (m, 1H) 2.22 (m, 2H) 2.05 (m, 1H)1.75 (m, 3H). LCMS (ESI pos) m/c 237 (M+1).

Step C: Preparation of1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole:To a 500 ml flask with stir bar was added4-chloro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (10.0 g, 42.2 mmol),DMSO (176 ml), PdCl₂(PPh₃)₂ (6.2 g, 8.86 mmol), tricyclohexylphosphine(0.47 g, 1.69 mmol), bis(pinacolato)diboron (16.1 g, 63.4 mmol) andpotassium acetate (12.4 g, 0.127 mol). With stirring, the mixture washeated to 130° C. for 16 hours. The reaction was cooled to 25° C. andEtOAc (600 ml) was added and washed with water (2×250 ml). The organicswere dried with MgSO₄ and concentrated in vacuo to dryness. The crudeproduct was purified by SiO₂ plug (120 g), eluting with 10%EtOAc/Hexanes (1 L) and 30% EtOAc/Hexanes (1 L). The filtrate wasconcentrated in vacuo to give 13.9 g (100%) of1-(Tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazoleas a 20% (wt/wt) solution in ethyl acetate. ¹H NMR shows the presence ofabout 20% (wt/wt) bis(pinacolato)diboron. ¹H NMR (400 MHz, CDCl₃) δ 8.37(s, 1H), 7.62 (dd, J=14 Hz, 2 Hz 1H), 7.60 (dd, J=7 Hz, 1 Hz 1H), 7.31(dd, J=8 Hz, 7 Hz 1H) 5.65 (dd, J=9 Hz, 3 Hz 1H) 4.05 (m, 1H) 3.75 (m,1H) 2.59 (m, 1H) 2.15 (m, 1H) 2.05 (m, 1H) 1.75 (m, 3H) 1.34 (s, 12H).LCMS (ESI pos) m/e 245 (M+1).

Example 5 1-(Tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)-1H-indazole (Route B)

Step A: Preparation of 4-nitro-1H-indazole: A mixture of2-methyl-3-nitro aniline (200 g, 1.315 moles), acetic acid (8000 ml) wascooled to 15-20° C. and a solution of sodium nitrite (90.6 g, 1.315moles) in water (200 ml) was slowly added over 30 min. After theaddition, the reaction temp. was increased to 25-30° C. and the reactionwas stirred at this temp for 2-3 h. Reaction progress was monitored byTLC and after completion of reaction product was filtered and residuewas washed with acetic acid (1000 ml). Acetic acid was distilled undervacuum (550 mm of Hg) below 80° C. and water (8000 ml) was added, cooledto 25-30° C. and stirred for 30 min. The slurry was filtered and washedwith water (1000 ml). Crude product was dried under heating at 70-80° C.for 2 hours, then was taken in 5% ethyl acetate/n-hexane (100:2000 ml)solution and stirred for 1-1.5 h at ambient temperature. The suspensionwas filtered and washed with 5% ethyl acetate/n-hexane mixture (25:475ml). The product obtained was dried under vacuum at below 80° C. for10-12 h to give 4-nitro-1H-indazole as a brown solid (150 g, 70%): mp:200-203° C.; ¹H NMR (200 MHz, CDCl₃) δ 13.4 (br, 1H), 8.6 (s, 1H),8.2-7.95 (dd, 2H), 7.4 (m, 1H). ESMS m/z 164 (M+1). Purity: 95% (HPLC)

Step B: Preparation of 4-amino-1H-indazole: A mixture of4-nitro-1H-indazole (200 g, 1.22 moles) and 10% palladium on carbon(20.0 g,) in EtOH (3000 ml) was hydrogenated at ambient temperature(reaction was exothermic and temperature increased to 50° C.). Aftercompletion of reaction, the catalyst was removed by filtration. Thesolvent was evaporated under vacuum at below 80° C. and cooled to roomtemperature and n-hexane (1000 ml) was added to the residue and stirredfor 30 min. Isolated solid was filtered and washed with n-hexane (200ml). Product was dried under vacuum at 70-80° C. for 10-12 h to give4-amino-1H-indazole as a brown solid (114 g, 70%), m. p.: 136-143° C. ¹HNMR (200 MHz, CDCl₃) δ 12 (br, 1H), 8.0 (s, 1H), 7.1-7.0 (dd, 2H), 6.5(d, 1H), 3.9 (m, 2H). ESMS m/z 134 (M+1). Purity: 90-95% (HPLC)

Step C: Preparation of 4-iodo-1H-indazole: A mixture of4-amino-1H-indazole (50.0 g, 0.375 moles) in water (100 ml) and con.hydrochloric acid (182 ml) was cooled to −10° C. To this a solution ofsodium nitrite (51.7 g, 0.75 moles) in water (75 ml) was added drop wiseat −10° C. in about 30-60 min. (during addition frothing was observed).In another flask a mixture of potassium iodide (311 g, 1.87 moles) inwater (3000 ml) was prepared at room temperature and to this abovecooled diazonium salt at 30-40° C. was added in about 30-40 min. Thereaction was maintained at 30° C. for 1 h and after completion ofreaction, ethyl acetate (500 ml) was added and the reaction mixture wasfiltered through Celite. The layers were separated and the aq. layer wasextracted with ethyl acetate (2×500 ml). The combined organic layerswere washed with 5% hypo solution (2×500 ml), brine (500 ml), dried(Na₂SO₄) and concentrated. Crude product was purified by chromatography(silica gel, hexane, 15-20% ethyl acetate/hexane) to furnish4-iodo-1H-indazole as an orange solid (23.0 g, 25%). mp: 151-177 C: ¹HNMR (200 MHz, CDCl₃) δ 12.4 (br, 1H), 8.0 (s, 1H), 7.6 (dd, 2H), 7.1 (d,1H). ESMS m/z 245 (M+1). Purity: 95-98% (HPLC).

Step D: Preparation of 4-iodo-1-(2-tetrahydropyranyl) indazole: Amixture of 4-amino-1H-indazole (250.0 g, 1.024 moles),3,4-dihydro-2H-pyran (126.0 g, 1.5 moles) and PPTS (2.57 g, 0.01 moles)in CH₂Cl₂ (1250 ml) was heated to 50° C. for 2 h. The reaction wascooled to room temperature and poured into water (625 ml), the layerswere separated, and aqueous layer was extracted with CH₂Cl₂ (250 ml).The combined organic layers were washed with water (625 ml), dried(Na₂SO₄) and concentrated. Crude residue was purified by chromatography(silica gel, hexane, 5-10% ethyl acetate/hexane) to furnish4-iodo-1-(2-tetrahydropyranyl) indazole as an oil (807.0 g, 60%). ¹H NMR(200 MHz, CDCl₃) δ 8.5 (s, 1H), 7.8 (m, 1H), 7.6 (d, 1H), 7.25 (m, 1H),5.7 (dd, 1H), 4.2-3.8 (dd, 1H), 2.2-2.0 (m, 4H) 2.0-1.8 (m, 4H). ESMSm/z 329 (M+1).

Step E: Preparation of1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole:A mixture of 4-iodo-1-(2-tetrahydropyranyl) indazole (100 g, 0.304moles), bispinacalotodiborane (96.4 g, 0.381 moles), PdCl₂ (dppf) (8.91g, 0.012 moles) and potassium acetate (85.97 g, 0.905 moles) in DMSO(500 ml) were heated to 80° C. for 2-3 h. After completion, reaction wascooled to room temperature and water (1500 ml) was added. Reaction masswas extracted into ethyl acetate (3×200 ml) and combined organic layerswere evaporated, dried (Na₂SO₄) and concentrated. Crude product waspurified by column chromatography (silica gel, hexane, 5-10% ethylacetate/hexane) to obtain1-(Tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazoleas viscous brown oil (70.0 g, 70%). ¹H NMR (CDCl₃) δ 8.5 (s, 1H), 7.8(m, 1H), 7.6 (d, 1H), 7.25 (m, 1H), 5.7 (dd, 1H), 4.2-3.8 (dd, 1H),2.2-2.0 (m, 4H) 2.0-1.8 (m, 4H) 1.4-1.2 (s, 12H). ESMS m/z 329 (M+1)

Example 6 4-methyl-5-(4,4,5,5-tetramethyl (1,3,2-dioxaborolan-2-yl))pyrimidine-2-ylamine

To a solution of 4-methylpyrimidine-2-ylamine (8.0 g, 0.073 mol) inchloroform (320 mL) was added N-bromosuccinimide (13.7 g, 0.077 mol).The reaction mixture was stirred in the dark for 18 hrs. LC/MS indicatedthe reaction was completed. The mixture was diluted with DCM, thenwashed with 1N NaOH aq solution and brine, dried over MgSO₄, filteredand concentrated to yield 5-bromo-4-methylpyrimidine-2-ylamin (12 g,Yield: 86%).

A mixture of 5-bromo-4-methylpyrimidine-2-ylamine (5.0 g, 26 mmol),potassium acetate (7.83 g, 79.8 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(7.43 g, 29.2 mmol) in dioxane (140 mL) was stirred for 20 min undernitrogen. 1, 1′-bis (diphenylphosphino) ferrocene palladium (II)chloride dichloromethane adduct (1.08 g, 1.33 mmol) was added to thereaction mixture. The reaction mixture was heated to 115° C. for 18 hunder nitrogen. Upon completion, the mixture was cooled and EtOAc wasadded. The resulting mixture was sonicated and filtered. AdditionalEtOAc was used to wash the solid. The combined organic extracts werewashed with water, dried over MgSO₄, filtered and concentrated. Thecrude was purified by chromatography eluting with 20-100% EtOAc/hexaneto yield 4.5 g of 4-methyl-5-(4,4,5,5-tetramethyl(1,3,2-dioxaborolan-2-yl))pyrimidine-2-ylamine (yield: 74%). ¹H-NMR(DMSO, 400 MHz): δ 8.28 (s, 1H), 6.86 (br s, 2H), 2.35 (s, 3H), 1.25 (s,12H). MS (ESI) m/e (M+H⁺) 236.15, 154.07.

Example 7 2,4,6-Trichloro-5-hydroxy-pyrimidine

Step 1: 6-Hydroxy-5-methoxy-1H-pyrimidine-2,4-dione, sodium salt

Under an atmosphere of nitrogen sodium metal (1.15 g, 0.05 moles) wasadded portionwise to ethanol (dried over 4 Å molecular sieves, 50 mL) at40° C. and the mixture stirred until a solution was formed. Urea (3.0 g,0.05 moles) was added and the mixture heated at 100° C. for 15 minutesuntil complete dissolution was achieved. The reaction mixture wasallowed to cool slightly and methoxymethylmalonate (8.1 g, 0.05 moles)added, a pink/white precipitate forming almost immediately. Further dryethanol (10 mL) was added to maintain a stirrable mixture. The resultantsuspension was heated at 100° C. (reflux) for 4 hours. The reactionmixture was concentrated in vacuo and the residue dried under highvacuum to give 6-Hydroxy-5-methoxy-1H-pyrimidine-2,4-dione, sodium saltas a pink/white solid used in the subsequent step without analysis orpurification.

Step 2: 2,4,6-Trichloro-5-methoxy-pyrimidine

6-Hydroxy-5-methoxy-1H-pyrimidine-2,4-dione, sodium salt (21 mmol) wassuspended in phosphorous oxychloride (20 mL) and the mixture divedbetween two 20 mL microwave reaction vials. The reaction mixtures wereheated at 130-140° C. (˜10-12 bar) for 30 minutes using microwaveirradiation (CARE! SIGNIFICANT PRESSURE INCREASE!) The cooled reactionmixtures were combined and poured onto warm (approximately 40° C.) water(CARE!) and the resultant mixture extracted twice with ethyl acetate,the combined organic extracts were dried (Na₂SO₄), filtered andconcentrated in vacuo to give 2,4,6-Trichloro-5-methoxy-pyrimidine as acrystalline yellow/brown solid (3.75 g, 84%). ¹H NMR (CDCl₃): 3.98 (3H,s).

Step 3: Under an atmosphere of nitrogen a solution of2,4,6-trichloro-5-methoxy-pyrimidine (4.0 g, 18.7 mmol) in DCM (200 mL)was cooled to 0° C. and treated with boron tribromide (neat, 6.6 mL, 65mmol) dropwise. After stirring for 18 hours at room temperature LCMSanalysis indicated complete reaction. The reaction mixture was cooledand diluted with methanol (25 mL, CARE, EXOTHERM!) and the reactionmixture diluted with water (200 mL). The aqueous layer was washed withDCM and the combined organic extracts dried (Na₂SO₄) filtered andconcentrated in vacuo to give 2,4,6-trichloro-5-hydroxy-pyrimidine as apale tan solid (2.55 g, 71%). ¹³C NMR (DMSO-d₆): 149.23 (C), 145.25 (C),145.08 (C). LCMS: R_(T)=2.65/2.77 [M−H]⁺=197/199.

Example 1011-[4-(3a,8-dimethyl-7-morpholin-4-yl-3,3a,8,8a-tetrahydro-2h-1-oxa-4,6,8-triaza-cyclopenta[a]inden-5-yl)-phenyl]-3-ethyl-urea101

Step 1: A mixture of 2,4-dichloro-5H-pyrrolo[3,2-d]pyrimidine (2.67 g,14.2 mmol), methanesulfonic acid methyl ester (1.26 mL, 14.9 mmol) andcesium carbonate (9.25 g, 28.4 mmol) in anhydrous N,N-dimethylformamide(21 mL) was stirred at RT (room temperature) under N₂ for 15 h. Thereaction mixture was diluted with ethyl acetate, and the organic layerwas washed with 1:1 water/brine (3×) and brine (1×), dried over Na₂SO₄,filtered, and concentrated in vacuo. The crude product was purified bysilica gel chromatography to give 2.50 g (87.1%) of2,4-dichloro-5-methyl-5H-pyrrolo[3,2-d]pyrimidine as a white solid.MS(ESI) m/z: 202.2/204.1 [M+1]⁺.

Step 2: A mixture of 2,4-dichloro-5-methyl-5H-pyrrolo[3,2-d]pyrimidine(1.25 g, 6.19 mmol), morpholine (1.08 mL, 12.37 mmol), andN,N-diisopropylethylamine (2.37 mL, 13.61 mmol) in N,N-dimethylformamide(36 mL) was stirred at RT under N₂ for 15 h. The reaction mixture wasdiluted with 1:1 diethyl ether/ethyl acetate, and the organic layer waswashed with 1:1 water/brine (3×) and brine (IX), dried over Na₂SO₄,filtered, and concentrated in vacuo. The crude product was purified bysilica gel chromatography to give 1.47 g (94.0%) of4-(2-chloro-5-methyl-5H-pyrrolo[3,2-d]pyrimidin-4-yl)morpholine as awhite solid. MS(ESI) m/z: 253.1/255.1 [M+1]⁺.

Step 3 To a stirred mixture of4-(2-chloro-5-methyl-5H-pyrrolo[3,2-d]pyrimidin-4-yl)morpholine (1.46 g,5.78 mmol) in tert-butyl alcohol (50 mL) and water (20 mL) at RT wasadded NBS (3.08 g, 17.33 mmol). The reaction mixture was stirred at 30°C. for 18 h. The reaction mixture was diluted with ethyl acetate, andthe organic layer was washed with water and brine, dried over Na₂SO₄,filtered, and concentrated in vacuo. The crude product was purified bysilica gel chromatography to give 2.45 g (99.4%) of7,7-dibromo-2-chloro-5-methyl-4-morpholino-5H-pyrrolo[3,2-d]pyrimidin-6(7H)-oneas a solid. MS(ESI) m/z: 427 [M+1]⁺.

Step 4: To7,7-dibromo-2-chloro-5-methyl-4-morpholino-5H-pyrrolo[3,2-d]pyrimidin-6(7H)-one(1.70 g, 3.99 mmol) in 2M aqueous ammonium chloride solution (9.96 mL,19.9 mmol) and THF (40 mL) at 0° C. was added zinc dust (573.4 mg, 8.775mmol). The reaction mixture was stirred at RT for 30 min and thendiluted with DCM (40 mL). The reaction mixture was filtered through apad of Celite. The layers were separated, and the organic layer waswashed with saturated aqueous NaHCO₃ solution, water, and brine, driedover Na₂SO₄, filtered, and concentrated in vacuo. The crude product waspurified by silica gel chromatography to give 679 mg (63.4%) of2-chloro-5-methyl-4-morpholino-5H-pyrrolo[3,2-d]pyrimidin-6(7H)-one asolid. MS(ESI) m/z: 269.2 [M+1]⁺.

Step 5: To2-chloro-5-methyl-4-morpholino-5H-pyrrolo[3,2-d]pyrimidin-6(7H)-one(466.0 mg, 1.73 mmol) in anhydrous THF (25 mL) under N₂ at −78° C. wasadded dropwise 1.0 M lithium hexamethyldisilazide in THF (3.8 mL, 3.8mmol). The reaction mixture was stirred at −78° C. under N₂ for 60minutes. Methyl iodide (0.32 mL, 5.20 mmol) was then added, and thereaction mixture was stirred at ambient temperature for 18 h. Thereaction mixture was quenched with saturated aqueous NH₄Cl solution anddiluted with ethyl acetate. The organic layer was washed with water andbrine, dried over Na₂SO₄, filtered, and concentrated in vacuo.Purification by silica gel chromatography eluted with 20 to 100% ethylacetate/heptane yielded two compounds:2-chloro-5,7,7-trimethyl-4-morpholino-5H-pyrrolo[3,2-d]pyrimidin-6(7H)-one(169.0 mg, 32.8%) as the first eluant; MS(ESI) m/z: 297.0 [M+1]⁺, and2-chloro-5,7-dimethyl-4-morpholino-5H-pyrrolo[3,2-d]pyrimidin-6(7H)-oneas second eluant (63.8 mg, 13.0%); MS(ESI) m/z: 283.2 [M+1]⁺.

Step 6: To2-chloro-5,7-dimethyl-4-morpholino-5H-pyrrolo[3,2-d]pyrimidin-6(7H)-one(92.0 mg, 0.32 mmol) in anhydrous THF (5 mL) under N₂ at −78° C. wasadded dropwise 1.0 M lithium hexamethyldisilazide in THF (0.65 mL, 0.65mmol). The reaction mixture was stirred at −78° C. under N₂ for 60minutes. Allyl bromide (0.062 mL, 0.72 mmol) dissolved in 0.5 mL of THFwas added, and the reaction mixture was stirred at ambient temperaturefor 16 h. The reaction mixture was quenched with saturated aqueous NH₄Clsolution and diluted with ethyl acetate. The organic layer was washedwith water and brine, dried over Na₂SO₄, filtered, and concentrated invacuo. Purification by silica gel chromatography eluted with 10 to 100%ethyl acetate/heptane gave 84.3 mg (80.3%) of7-allyl-2-chloro-5,7-dimethyl-4-morpholino-5H-pyrrolo[3,2-d]pyrimidin-6(7H)-one.MS(ESI) mm/z: 323.1 [M+1]⁺.

Step 7: To a stirred solution of7-allyl-2-chloro-5,7-dimethyl-4-morpholino-5H-pyrrolo[3,2-d]pyrimidin-6(7H)-one(80.0 mg, 0.25 mmol) in anhydrous THF (3.0 m) and water (1.0 mL) cooledto 0° C. was added N-methylmorpholine N-oxide (34.8 mg, 0.29 mmol),followed by 2.5% osmium tetraoxide in tert-butanol. (0.033 mL, 0.025mmol). The reaction mixture was stirred at ambient temperature under N₂for 16 h. Sodium sulfite (312.4 mg, 2.48 mmol) was then added, and thereaction mixture was stirred at RT for 20 min. The reaction mixture wasdiluted with water and then extracted with ethyl acetate (2×). Thecombined organic layers were dried over Na₂SO₄, filtered, andconcentrated in vacuo. The crude intermediate was then diluted withanhydrous THF (3.0 mL) and water (1.0). Sodium periodate (79.5 mg, 0.372mmol) was added, and the reaction mixture was sonicated for 1 minute andthen stirred at RT for 2 days. The reaction mixture was diluted withethyl acetate, and the organic layer was washed with water and brine,dried over Na₂SO₄, filtered, and concentrated in vacuo to give 80.4 mg(99%) of2-(2-chloro-5,7-dimethyl-4-morpholino-6-oxo-6,7-dihydro-5H-pyrrolo[3,2-d]pyrimidin-7-yl)acetaldehyde.MS(ESI) m/z: 325.1 [M+1]⁺.

Step 8: To a stirred solution of2-(2-chloro-5,7-dimethyl-4-morpholino-6-oxo-6,7-dihydro-5H-pyrrolo[3,2-/d]pyrimidin-7-yl)acetaldehyde(80.4 mg, 0.248 mmol) in THF (3.7 mL) and methanol (0.25 mL) at 0° C.was added sodium borohydride (20.6 mg, 0.55 mmol) in one portion. Thereaction mixture was stirred at ambient temperature under N₂ for 5 h andthen diluted with ethyl acetate. The organic layer was washed with waterand brine, dried over Na₂SO₄, filtered, and concentrated in vacuo.Purification by silica gel chromatography eluted with 15 to 100% ethylacetate/heptane gave 76 mg (99%) of4-(2-chloro-5-methyl-{3a,6-dimethylhexahydro-2H-furo[2,3-b]pyrrolo}[3,2-d]pyrimidin-4-yl)morpholine.MS(ESI) m/z: 311.2 [M+1]⁺.

Step 9: In a microwave vial was placed4-(2-chloro-5-methyl-{3a,6-dimethylhexahydro-2H-furo[2,3-b]pyrrolo}[3,2-d]pyrimidin-4-yl)morpholine(70.0 mg, 0.225 mmol), 4-nitrophenylboronic acid pinacol ester (70.1 mg,0.281 mmol), tetrakis(triphenylphosphine)palladium(0) (18.2 mg, 0.016mmol), sodium carbonate (41.1 mg, 0.38 mmol), and potassium carbonate(49.8 mg, 0.36 mmol). Degassed acetonitrile (3.5 mL) and degassed water(1.0) were added. The reaction mixture was subjected to microwaveirradiation at 120° C. for 15 minutes. The cooled reaction was dilutedwith ethyl acetate, and the reaction mixture was filtered through a padof Celite to rid excess Pd. The organic layer was washed with water andbrine, dried over Na₂SO₄, filtered, and concentrated in vacuo.Purification by silica gel chromatography eluted with 10 to 100% ethylacetate/heptane gave 75.3 mg (84.1%) of4-(5-methyl-2-(4-nitrophenyl)-{3a,6-dimethylhexahydro-2H-furo[2,3-h]pyrrolo}[3,2-d]pyrimidin-4-yl)morpholine.MS(ESI) m/z: 398.3 [M+1]⁺.

Step 10: To a stirred solution of4-(5-methyl-2-(4-nitrophenyl)-{3a,6-dimethylhexahydro-2H-furo[2,3-b]pyrrolo}[3,2-d]pyrimidin-4-yl)morpholine(78.0 mg, 0.196 mmol) dissolved in ethanol (4.7 mL) and water (3.1 mL)was added ammonium chloride (210 mg, 3.93 mmol) followed by iron (54.8mg, 0.982 mmol). The reaction mixture was stirred at 95° C. for 2 h andthen diluted with 10% MeOH/DCM and saturated aqueous NaHCO₃ solution.The reaction mixture was sonicated and then filtered through a pad ofCelite to rid iron. The filtrate was extracted with DCM (3×), and thecombined organic layers were washed with brine, dried over Na₂SO₄,filtered, and concentrated in vacuo to obtain 68.0 mg (94.2%) of4-(5-methyl-4-morpholino-{3a,6-dimethylhexahydro-2H-furo[2,3-b]pyrrolo}[3,2-d]pyrimidin-2-yl)anilineas a white foam. MS(ESI) nm/z: 368.2 [M+1]⁺.

Step 11: To a stirred solution of4-(5-methyl-4-morpholino-{3a,6-dimethylhexahydro-2H-furo[2,3-b]pyrrolo}[3,2-d]pyrimidin-2-yl)aniline(17.4 mg, 0.047 mmol) in anhydrous 1,2-dichloroethane (1.4 mL) was addedethyl isocyanate (0.037 mL, 0.47 mmol), and the reaction mixture wasstirred at 50° C. under N₂ for 2 h. The reaction was quenched with MeOH(1 mL). Volatile solvent was removed under reduced pressure, and thecrude was purified by HPLC to give 11.50 mg (55.4%) of 101 as fluffywhite solid. ¹H NMR (400 MI-Hz, DMSO) e 8.57 (s, 1H), 8.14 (d, J=8.8 Hz,2H), 7.45 (d, J=8.8 Hz, 2H), 6.12 (t, J=5.6 Hz, 1H), 4.98 (s, 1H), 3.87(t, J=7.0 Hz, 1H), 3.84-3.66 (m, 4H), 3.60-3.48 (m, 2H), 3.43-3.36 (m,1H), 3.34-3.28 (m, 2H), 3.16-3.06 (m, 2H), 2.81 (s, 3H), 2.15 (dd,J=12.2, 3.4 Hz, 1H), 2.03-1.88 (m, 1H), 1.47 (s, 3H), 1.06 (t, J=7.2 Hz,3H). MS(ESI) m/z: 439.2 [M+1]⁺.

Example 1025-(6,6-dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)-4-methylpyrimidin-2-amine102

A mixture of2-(2-(2-amino-4-methylpyrimidin-5-yl)-9-(2-hydroxyethyl)-6-morpholino-9H-purin-8-yl)propan-2-ol(550 mg, 1.3 mmol) and TFA (0.36 mL, 4.7 mmol) in toluene (9 mL) washeated at 110° C. and stirred 4 h. The reaction mixture was cooled toroom temperature and concentrated in vacuo. Analytical LC-MS indicatedconversion to the cyclic product as well as the elimination by-product,2-(2-(2-amino-4-methylpyrimidin-5-yl)-6-morpholino-8-(prop-1-en-2-yl)-9H-purin-9-yl)ethanol.The crude residue was dissolved in DMF (1 mL) and purified bypreparative rp-HPLC. This process provided 302 mg (57% yield) of 102. MS(ESI+): m/z 397.4 (M+H⁺). ¹H NMR (400 MHz, DMSO) δ 8.79 (s, 1H), 6.78(s, 2H), 4.20 (s, 4H), 4.12 (s, 4H), 3.74 (s, 5H), 2.63 (s, 4H), 1.58(s, 7H)

Example 1035-(6,6-dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine103 Step 1:2-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)propan-2-ol

4-(2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine (20.0g, 0.062 mol) was cooled to −42° C. in THF (400 mL). A solution ofn-butyllithium (2.5 M in hexanes, 48 mL, 0.12 mol) was added portionwiseover 10 min. The mixture gradually turned yellow. The reaction mixturewas then allowed to stir at −42° C. for 30 minutes and then anhydrousacetone (10 mL, 0.1 mol) was added at once. The resulting reactionmixture was slowly warmed to 0° C. over a 2 hour period. The mixture wassubsequently quenched with water, extracted with EtOAc and dried overMgSO₄. The slurry was filtered and concentrated in vacuo to provide2-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)propan-2-ol(23 g, 98%) as a yellow solid. MS (ESI+): m/z 382.1(M+H⁺)₂-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)propan-2-ol

Step 2:2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine

2-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)propan-2-ol(23 g, 0.06 mol) was suspended in MeOH (270 mL) and a catalytic amountof p-toluenesulfonic acid monohydrate (1.22 g, 7.1 mmol) was added. Thereaction mixture was heated to 50° C. and stirred for 16 h. The solutionbecame homogenous with extended heating. LC-MS indicated completeconversion to the THP-deprotected product. The reaction mixture wasconcentrated to completely remove MeOH and the resultant solid wassubsequently diluted with water and EtOAc. The phases were partitionedand the aqueous was extracted three times with EtOAc, dried over MgSO₄,filtered and concentrated. Crude2-(2-chloro-6-morpholino-9H-purin-8-yl)propan-2-ol (ca. 15.0 g, 50.4mmol) was treated with 1,2-dibromoethane (8.7 mL, 100 mmol) and cesiumcarbonate (41.0 g, 126 mmol) in DMF (200 mL). The reaction mixture washeated at 90° C. for 1.5 h. LC-MS indicated complete conversion to thecyclic product with ˜10% of E2 elimination product present. The reactionmixture was cooled to room temperature and poured into a separatoryfunnel containing 1 N HCl and EtOAc (50:50). The aqueous layer wasextracted several times with EtOAc and the combined organic portionswere washed once with water. Subsequent drying over MgSO₄ was followedby filtration and concentration to yield a crude oily residue. Thismaterial was loaded onto a 300-g ISCO column and purified by slowgradient flash column chromatography (15-30% EtOAc in heptane).Fractions containing the desired product were concentrated and drieddown under high vacuum pressure for an overnight period to give 14.3 g(88% yield) of2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine.MS (ESI+): m/z 324.2 (M+H⁺). ¹H NMR (400 MHz, DMSO) δ 4.07 (m, 8H), 3.72(m, 4H), 1.57 (s, 6H)

Step 3:2-Chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(180 mg, 0.56 mmol) in 1,2-dimethoxyethane (5.1 mL) was added5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine 25 (180mg, 0.83 mmol) and 1.0 M of cesium carbonate in water (1.7 mL). Thereaction mixture was degassed for 5 min and recycled with nitrogenatmosphere. Subsequently,1,1′-bis(diphenylphosphino)ferrocenepalladium(II) chloride (54 mg, 0.067mmol) was added, and the mixture was degassed and recycled again. Thereaction vial was then subjected to microwave irradiation for 20 mins at140° C. Solid precipitate that formed during the reaction was filteredand rinsed with excess water. The precipitate was taken up in DCM andpurified by FCC (40 g, 0.5-4% MeOH in DCM, slow gradient) to isolate 103as a tan powder (56 mg, 27% yield). MS (ESI+): m/z 383.1 (M+H⁺). ¹H NMR(400 MHz, DMSO) δ 9.09 (s, 2H), 7.00 (s, 2H), 4.23 (m, 4H), 4.13 (m,4H), 3.79-3.68 (m, 4H), 2.50 (s, 6H)

Example 1045-(6,6-dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)-4-(trifluoromethyl)pyridyl-2-amine104

2-Chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(180 mg, 0.56 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)romoace-2-amine (240 mg, 0.83 mmol) were reacted under microwave Suzukipalladium conditions to give 104 (204 mg, 70% yield). LC/MS (ESI+): m/z450 (M+H). ¹H NMR (400 MHz, DMSO) δ 8.47 (s, 1H), 6.82 (s, 1H), 6.70 (s,2H), 4.19 (s, 4H), 4.11 (s, 4H), 3.71 (s, 4H), 1.59 (s, 6H)

Example 1055-(4-morpholino-8,9-dihydro-7h-[1,3]oxazino[2,3-e]purin-2-yl)pyrimidin-2-amine105 Step 1:4-(2-chloro-8-iodo-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine

To a solution of4-(2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine (1.0g, 3.1 mmol) and N,N,N′,N′-tetramethylenediamine (0.7 mL, 4.6 mmol) inanhydrous THF (23 mL) at −42° C. was added 2.5 M n-butyllithium solutionin hexane (4.3 mL, 11.0 mmol) dropwise down the side of the reactionflask. The reaction was stirred at cold temperature and maintained for 1h before 1-chloro-2-iodoethane (1.4 mL, 15 mmol) was introduced.Stirring was continued for 1.5 h period. LC-MS indicated the reactionhad reached complete conversion to the desired product. The reactionmixture was subsequently quenched and worked up with sat. NH₄Cl aqueoussolution which was extracted with EtOAc (3 times), dried over MgSO₄,filtered and concentrated in vacuo to give4-(2-chloro-8-iodo-9-(tetrahydro-21H-pyran-2-yl)-9H-purin-6-yl)morpholine(1.4 g, 84% yield) determined by LC-MS to be >90% purity. MS (ESI+): m/z450.1 (M+H⁺)

Step 2:4-(2-chloro-8-iodo-9-(3-(tetrahydro-2H-pyran-2-yloxy)propyl)-9H-purin-6-yl)morpholine

To a suspension of4-(2-chloro-8-iodo-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine(0.655 g, 1.46 mmol) in methanol (6 mL) was added a catalytic amount ofp-toluenesulfonic acid (25 mg, 0.14 mmol). The reaction mixture washeated at 50° C. for an overnight period. After this time, the mixturewas cooled to room temperature and the volume of methanol was reduced byvacuum evaporation. The resulting residue was diluted with sat. aqueousNaHCO₃ solution. The precipitate that formed was collected byfiltration. In total, 295 mg (56%) of4-(2-chloro-8-iodo-9H-purin-6-yl)morpholine was obtained as a whitesolid which was dissolved in anhydrous DMF (2.5 mL). Cesium carbonate(0.53 g, 1.61 mmol) was added and the mixture was stirred together 10min at 23° C. Subsequently1-(2H-3,4,5,6-tetrahydropyran-2-yloxy)-3-bromopropane (0.54 g, 2.42mmol) was introduced to the mixture. The resulting reaction mixture washeated at 50° C. for 2 h. Complete conversion was observed at the end ofthis period. The reaction was worked up by dilution with 1 N HCl andEtOAc. The phases were separated and the aqueous layer was extractedtwice with EtOAc. The combined organic portions were dried over MgSO₄,filtered and concentrated in vacuo. The residue was purified by FCC (40g silica gel column, 0-50% EtOAc in heptane) to give 385 mg (94% yield)of4-(2-chloro-8-iodo-9-(3-(tetrahydro-2H-pyran-2-yloxy)propyl)-9H-purin-6-yl)morpholineas a pale yellow solid. MS (ESI+): m/z 508.0 (M+H⁺)

Step 3: 2-chloro-4-morpholino-8,9-dihydro-7H-[1,3]oxazino[2,3-e]purine

To4-(2-chloro-8-iodo-9-(3-(tetrahydro-2H-pyran-2-yloxy)propyl)-9H-purin-6-yl)morpholine(0.39 g, 0.8 mmol) in methanol (5 mL) was added p-toluenesulfonic acid(10 mg, 0.08 mmol). The reaction was heated at 50° C. for 30 min,whereupon precipitation was observed signaling reaction completion. Thiswas confirmed by analytical LC-MS. The reaction mixture was concentratedin vacuo to give3-(2-chloro-8-iodo-6-morpholino-9H-purin-9-yl)propan-1-ol. Copper(I)iodide (9 mg, 0.05 mmol), picolinic acid (10 mg, 0.09 mmol) andpotassium phosphate (0.4 g, 1.9 mmol) were combined in oven-dried 50-mLround-bottom flask and evacuated/recycled with N₂ atmosphere.Subsequently, 3-(2-chloro-8-iodo-6-morpholino-9H-purin-9-yl)propan-1-olwas dissolved in anhydrous dimethyl sulfoxide (6.7 mL) and introducedvia syringe. The entire mixture was heated at 80° C. for 20 h. LC-MSindicated good conversion to the desired product. The reaction mixturewas cooled to room temperature, diluted with water and EtOAc, dried overMgSO4, filtered and concentrated in vacuo. The residue was purified byFCC (40 g silica gel column, 0-100% EtOAc in heptane) to give 112 mg of2-chloro-4-morpholino-8,9-dihydro-7H-[1,3]oxazino[2,3-e]purine (40%). MS(ESI+): m/z 296.2 (M+H⁺)

Step 4: 2-chloro-4-morpholino-8,9-dihydro-7H-[1,3]oxazino[2,3-e]purine(112 mg, 0.38 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine 25 (0.1g, 0.454 mmol), 1 M cesium carbonate in water (0.8 mL, 0.8 mmol) andacetonitrile (0.8 mL) were placed in a 10-mL CEM microwave reaction vialand capped. The flask was slowly evacuated under vacuum and replacedwith a nitrogen atmosphere. Tetrakis(triphenylphosphine)palladium(0)(43.8 mg, 0.038 mmol) was introduced and the evacuation/N₂ cycle wasrepeated. The reaction mixture was irradiated in the microwave at 140°C. for 20 min. LC-MS indicated complete conversion. The mixture wasfiltered through a short plug of Celite® eluting with EtOAc andsubsequently concentrated in vacuo. The residue was purified by rp-HPLCto provide 105 (62.3 mg, 46% yield). MS (ESI+): m/z 355.1 (M+H⁺)

Example 1065-(4-morpholino-6,7,8,9-tetrahydropyrido[2,1-e]purin-2-yl)pyrimidin-2-amine106 Step 1:4-(8-(4-bromobutyl)-2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine

A solution of4-(2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine(Example 118) (5.0 g, 15 mmol) and N, N, N′,N′-tetramethylethylenediamine (3.5 mL, 23 mmol) in THF (110 mL) wascooled to −42° C. and treated with a solution of 2.5M n-Butyllithium inhexane (22 mL, 54 mmol) drop-wise over 5 minutes. After 30 minutes at−42° C., 1, 4-dibromo-butane (8.9 mL, 75 mmol) was added and thereaction mixture was slowly warmed to 0° C. over 1 hr and then warmed toambient temperature for 90 minutes. The mixture was quenched with asaturated solution of NH₄Cl, and diluted with ethyl acetate. The aqueouslayer was extracted into ethyl acetate (3×), and the combined organicswere dried over sodium sulfate, filtered, and absorbed onto celite forpurification by flash chromatography to afford4-(8-(4-bromobutyl)-2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholineas a white solid (1.1 g, 15%). LC/MS (ESI+): m/z 459 (M+H)

Step 2: 4-(8-(4-bromobutyl)-2-chloro-9H-purin-6-yl)morpholine

A suspension of4-(8-(4-bromobutyl)-2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine(1.1 g, 2.4 mmol) in methanol (10 mL) was treated with p-toluenesulfonicacid (40 mg, 0.24 mmol) and heated overnight at 50° C. The solvent wasremoved in vacuo to afford4-(8-(4-bromobutyl)-2-chloro-9H-purin-6-yl)morpholine as a white solid,which was used in the next step without any further purification (880mg, quant). LC/MS (ESI+): m/z 375 (M+H)

Step 3: 4-(2-chloro-6,7,8,9-tetrahydropyrido[2,1-e]purin-4-yl)morpholine

A suspension of 4-(8-(4-bromobutyl)-2-chloro-9H-purin-6-yl)morpholine(880 mg, 2.4 mmol) in DMSO (6.7 mL) was treated with cesium carbonate(1.5 g, 4.7 mmol) and heated at 50° C. for 1 hr. The reaction mixturewas cooled to ambient temperature and diluted with water and DCM and thelayers separated. The aqueous phase was extracted into DCM (3×), driedover sodium sulfate, filtered, and absorbed onto celite for purificationby flash chromatography to afford4-(2-chloro-6,7,8,9-tetrahydropyrido[2,1-e]purin-4-yl)morpholine as awhite solid (460 mg, 67%). ¹H NMR (400 MHz, DMSO) δ 4.28-4.03 (m, 4H),4.00 (m, 21H), 3.69 (m, 4H), 2.90 (m, 2H), 2.01-1.86 (m, 4H). LC/MS(ESI+): m/z 294 (M+H)

Step 4: Following General Procedure A,4-(2-chloro-6,7,8,9-tetrahydropyrido[2,1-e]purin-4-yl)morpholine (310mg, 1.0 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (300mg, 1.4 mmol), and tetrakis(triphenylphosphine)-palladium(0) (61 mg, 52umol, 5.0 mol %) suspended in MeCN (2.6 mL) and 1.0M Na₂CO₃ (2.0 mL) washeated under microwave irradiation at 140° C. for 15 minutes. The cooledreaction mixture was concentrated to dryness in vacuo. The resultantresidue was purified via HPLC to afford 106 as a white solid (220 mg,60%). ¹H NMR (500 MHz, DMSO) δ 9.09 (s, 2H), 7.01 (s, 2H), 4.23 (s, 4H),4.08 (s, 2H), 3.73 (s, 4H), 2.92 (s, 2H), 2.00 (s, 2H), 1.93 (s, 2H).LCMS: R_(T)=6.13 min, M+H⁺=353.1

Example 1075-(4-morpholino-6,7,8,9-tetrahydropyrido[2,1-e]purin-2-yl)pyridin-2-amine107

4-(2-chloro-6,7,8,9-tetrahydropyrido[2,1-e]purin-4-yl)morpholine, fromExample 106 (150 mg, 0.50 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (140 mg,0.64 mmol), and tetrakis(triphenylphosphine)-palladium(0) (29 mg, 25umol, 5.0 mol %) suspended in MeCN (1.2 mL) and 1.0M Na₂CO_(3(aq)) (0.94mL) were heated under microwave irradiation at 140° C. for 15 minutes.The cooled reaction mixture was concentrated to dryness in vacuo. Theresultant residue was purified via HPLC to afford 107 as a white solid(170 mg, 63%). ¹H NMR (500 MHz, DMSO) δ 8.91 (s, 1H), 8.27 (d, J=8.6 Hz,1H), 6.48 (d, J=8.5 Hz, 1H), 6.27 (s, 2H), 4.22 (s, 4H), 4.07 (s, 2H),3.73 (s, 4H), 2.91 (s, 2H), 2.01 (s, 2H), 1.92 (s, 2H). LCMS: R_(T)=6.23min, M+H⁺=352.1

Example 1085-(4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)-4-(trifluoromethyl)pyridyl-2-amine108

2-Chloro-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine (80 mg,0.0003 mol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)romoace-2-amine (120 mg, 0.0004 mol) were reacted under microwave Suzukipalladium conditions to give 108 (40 mg, 40% yield). LC/MS (ESI+): m/z422 (M+H). ¹H NMR (400 MHz, DMSO) δ 8.56-8.42 (m, 1H), 6.81 (s, 1H),6.76 (s, 2H), 4.93 (s, 2H), 4.16 (d, J=20.6 Hz, 8H), 3.80-3.58 (m, 4H)

Example 1095-(4-morpholino-7,8-dihydro-6h-pyrrolo[2,1-e]purin-2-yl)pyrimidin-2-amine109 Step 1: 4-(9-allyl-2-chloro-8-iodo-9H-purin-6-yl)morpholine

4-(2-chloro-8-iodo-9H-purin-6-yl)morpholine (500 mg, 1.0 mmol) wasstirred together with cesium carbonate (890 mg, 2.7 mmol) in DMF (4.2mL) at ambient temperature for 10 minutes. Allyl bromide (0.36 mL, 4.1mmol) was introduced and the reaction mixture was heated at 50° C. for 2hr. The mixture was cooled to ambient temperature and diluted with brineand DCM and the layers separated. The aqueous phase was extracted intoDCM (3×), dried over sodium sulfate, filtered, and absorbed onto celitefor purification by flash chromatography to afford4-(9-allyl-2-chloro-8-iodo-9H-purin-6-yl)morpholine as a white foam (480mg, 90%). ¹H NMR (500 MHz, DMSO) δ 5.94 (d, J=10.6 Hz, 2H), 5.76 (s,1H), 5.19 (d, J=10.3 Hz, 2H), 4.79 (d, J=16.9 Hz, 2H), 4.69 (s, 2H),3.73 (s, 4H). LC/MS (ESI+): m/z 406 (M+H)

Step 2: 4-(2-chloro-7,8-dihydro-6H-pyrrolo[2,1-e]purin-4-yl)morpholine

4-(9-Allyl-2-chloro-8-iodo-9H-purin-6-yl)morpholine (230 mg, 0.57 mmol)was added to a solution of 0.50M 9-borabicyclo[3.3.1]nonane in hexane(1.7 mL) at ambient temperature. THF was added to solubilize thereaction mixture. No conversion prompted addition of 0.50M9-borabicyclo[3.3.1]nonane in hexane (1.7 mL) and the reaction mixturestirred for 15 hr. Potassium phosphate monohydrate (200 mg, 0.85 mmol)and tetrakis(triphenylphosphine)-palladium(0) (16 mg, 14 umol, 2.5 mol%) were added and the reaction mixture was heated at 60° C. for 15 hr.The mixture was cooled to ambient temperature and diluted with water andDCM and the layers separated. The aqueous phase was extracted into DCM(3×), dried over sodium sulfate, filtered, and absorbed onto celite forpurification by flash chromatography to afford4-(2-chloro-7,8-dihydro-6H-pyrrolo[2,1-e]purin-4-yl)morpholine as asolid (32 mg, 20%). LC/MS (ESI+): m/z 280 (M+H)

Step 3: 4-(2-chloro-7,8-dihydro-6H-pyrrolo[2,1-e]purin-4-yl)morpholine(32 mg, 0.11 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (33 mg,0.15 mmol), and tetrakis(triphenylphosphine)-palladium(0) (6.6 mg, 5.7umol, 5.0 mol %) suspended in MeCN (0.28 mL) and 1.0M Na₂CO₃ (0.22 mL)was heated under microwave irradiation at 140° C. for 15 minutes. Thecooled reaction mixture was concentrated to dryness in vacuo. Theresultant residue was purified via HPLC to afford 109 as a white solid(1.7 mg, 4.4%). ¹H NMR (400 MHz, DMSO) δ 8.99 (s, 1H), 7.75 (s, 1H),6.95 (d, J=1.5 Hz, 1H), 6.92 (s, 1H), 6.88 (s, 1H), 6.02 (s, 1H), 5.85(hept, J=6.6 Hz, 1H), 4.65 (s, 1H), 4.50-4.36 (m, 6H), 3.73 (s, 6H),2.24 (s, 3H), 1.44 (d, J=6.6 Hz, 6H). LCMS: R_(T)=3.48 min, M+H⁺=339.1

Example 110 6,6-dimethyl-4-morpholino-2-(lH-pyrrolo[2,3-b]pyridin-5-yl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine110

2-Chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(100 mg, 0.0003 mol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrrolo[2,3-b]pyridine(110 mg, 0.00046 mol) were reacted under microwave Suzuki palladiumconditions to give 110 (54 mg, 50% yield). LC/MS (ESI+): m/z 406 (M+H).¹H NMR (400 MHz, DMSO) δ 11.75 (s, 1H), 9.27 (d, J=1.8 Hz, 1H), 8.88 (d,J=1.6 Hz, 1H), 7.60-7.41 (m, 1H), 6.57 (d, J=1.6 Hz, 1H), 4.29 (s, 4H),4.17 (dd, J=18.0, 5.1 Hz, 4H), 3.88-3.69 (m, 4H), 1.60 (s, 6H)

Example 1115-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyridin-2-amine11l

2-Chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(100 mg, 0.0003 mol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)romoace-2-amine (110 mg, 0.0005 mol) were reacted under microwave Suzukipalladium conditions to give 111 (75 mg, 70% yield). LC/MS (ESI+): m/z382 (M+H). ¹H NMR (400 MHz, DMSO) δ 8.92 (d, J=2.1 Hz, 1H), 8.27 (dd,J=8.7, 2.2 Hz, 1H), 8.16 (s, 1H), 6.49 (d, J=8.7 Hz, 1H), 6.26 (s, 2H),4.22 (s, 4H), 4.12 (s, 4H), 3.86-3.65 (m, 4H), 1.57 (s, 6H)

Example 1125-(4-morpholino-8,9-dihydrospiro[[1,3]oxazino[2,3-e]purine-7,1′-cyclopropane]-2-yl)pyrimidin-2-amine112 Step 1:1-(2-(2-chloro-6-morpholino-9H-purin-9-yl)ethyl)cyclopropanol

A solution of ethyl 3-(2-chloro-6-morpholino-9H-purin-9-yl)propanoate(500 mg, 1.5 mmol) in diethylether (37 mL) was treated with titanium(IV) ethoxide (31 uL, 0.15 mmol) followed by the dropwise addition of3.0M ethylmagnesium bromide in ether (0.98 mL, 2.9 mmol) at ambienttemperature for 90 minutes. Partial conversion prompted the addition oftitanium (IV) ethoxide (31 uL, 0.15 mmol) and 3.0M ethylmagnesiumbromide in ether (0.98 mL, 2.9 mmol). After 2 hr, the reaction mixturewas quenched with an aqueous 1.0M solution of HCl (20 mL) and filteredthrough a plug of celite, washing with ethyl acetate. The mixture wasdiluted with water and ethyl acetate and the layers separated. Theaqueous phase was extracted into EtOAc (3×), dried over sodium sulfate,filtered, and absorbed onto celite for purification by flashchromatography to afford1-(2-(2-chloro-6-morpholino-9H-purin-9-yl)ethyl)cyclopropanol as acolorless oil (220 mg, 46%). LC/MS (ESI+): m/z 324 (M+H)

Step 2:1-(2-(2-chloro-8-iodo-6-morpholino-9H-purin-9-yl)ethyl)cyclopropanol

A solution of1-(2-(2-chloro-6-morpholino-9H-purin-9-yl)ethyl)cyclopropanol (220 mg,0.68 mmol) and N,N,N′,N′-tetramethylethylene-diamine (0.15 mL, 1.0 mmol)in THF (4.9 mL) was cooled to −42° C. and treated with a solution of2.5M n-butyllithium in hexane (1.5 mL, 3.7 mmol) dropwise over 5minutes. After 30 minutes at −42° C., 1-chloro-2-iodoethane (0.31 mL,3.3 mmol) was added and the reaction mixture was slowly warmed to 0° C.over 1 hr. The mixture was quenched with a saturated solution of NH₄Cl,and diluted with ethyl acetate. The aqueous layer was extracted intoethyl acetate (3×), and the combined organics were dried over sodiumsulfate, filtered, and absorbed onto celite for purification by flashchromatography to afford1-(2-(2-chloro-8-iodo-6-morpholino-9H-purin-9-yl)ethyl)cyclopropanol asa colorless oil (220 mg, 71%). LC/MS (ESI+): m/z 450 (M+H)

Step 3:2-chloro-4-morpholino-8,9-dihydrospiro[[1,3]oxazino[2,3-e]purine-7,1′-cyclopropane]

Copper (I) iodide (4.6 mg, 24 umol), picolinic acid (6.0 mg, 48 umol),and potassium phosphate (210 mg, 0.97 mmol) were combined in anoven-dried round bottom flask and evacuated/recycled with N₂ (3×).Subsequently, a solution of1-(2-(2-chloro-8-iodo-6-morpholino-9H-purin-9-yl)ethyl)cyclopropanol(220 mg, 0.48 mmol) dissolved in DMSO (3.4 mL) was introduced viasyringe. The reaction mixture was heated at 80° C. for 20 hr. Thereaction mixture was cooled to ambient temperature and diluted withwater and ethyl acetate and the layers separated. The aqueous phase wasextracted into EtOAc (3×), dried over sodium sulfate, filtered, andabsorbed onto celite for purification by flash chromatography to afford2-chloro-4-morpholino-8,9-dihydrospiro[[1,3]oxazino[2,3-e]purine-7,1′-cyclopropane]as a yellow solid (41 mg, 26%). LC/MS (ESI+): m/z 322 (M+H)

Step 4:2-chloro-4-morpholino-8,9-dihydrospiro[[1,3]oxazino[2,3-e]purine-7,1′-cyclopropane](37 mg, 0.11 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (33 mg,0.15 mmol), and tetrakis(triphenylphosphine)-palladium(0) (6.6 mg, 5.7umol, 5.0 mol %) suspended in MeCN (0.28 mL) and 1.0M Na₂CO₃ (0.22 mL)was heated under microwave irradiation at 140° C. for 15 minutes. Thecooled reaction mixture was concentrated to dryness in vacuo. Theresultant residue was purified via HPLC to afford 112 as a white solid(22 mg, 50%). ¹H NMR (400 MHz, DMSO) δ 9.07 (s, 2H), 6.96 (s, 2H), 4.22(t, J=6.0 Hz, 2H), 4.11 (s, 4H), 3.75-3.65 (m, 4H), 2.26 (t, J=5.9 Hz,2H), 1.08 (t, J=6.4 Hz, 2H), 0.90 (t, J=6.6 Hz, 2H). LCMS: R_(T)=3.57min, M+H⁺=381.1.

Example 1135-(4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine113 Step 1:2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine-8-carbaldehyde

To a mixture of4-(2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine (5 g,20 mmol) in THF (100 mL) at −78° C. was added tetramethylethylenediamine(3.5 mL, 23 mmol) followed by 2.5 M of n-BuLi (9.3 mL, 23 mmol)dropwise. The reaction was stirred at −78° C. for 1 hour and thenN,N-dimethylformamide (2.4 mL, 31 mmol) was added, continue stirred for1 hours at −78° C. The reaction was quenched with water and extractedwith EtOAc. The organic extracts were washed with water, brine, driedover MgSO₄ and concentrated. The crude material was triturated withEtOAc to give pure2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine-8-carbaldehydeas a white solid (4.5 g, 80% yield). LC/MS (ESI+): m/z 353 (M+H)

Step 2:(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)methanol

2-Chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purine-8-carbaldehyde(3.8 g, 11 mmol) in MeOH (22 mL) was treated with sodiumtetrahydroborate (0.817 g, 22 mmol), and stirred at room temperature for1 hour. The reaction was quenched with water and extracted with EtOAc.The organic extracts were washed with water, brine, dried over MgSO₄ andconcentrated. The crude material was purified with isco with 0-80%EtOAc/hexane to give the pure(2-chloro-6-morpholino-9-(tetrahydro-2-pyran-2-yl)-9H-purin-8-yl)methanol(3.4 g, 89% yield). LC/MS (ESI+): m/z 354 (M+H)

Step 3: (2-chloro-6-morpholino-9H-purin-8-yl)methanol

(2-Chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)methanol(3.4 g, 0.0096 mol) in MeOH (20 mL) was treated with a catalytic amountof p-toluenesulfonic acid (0.25 g, 0.00144 mol). The reaction mixturewas heated to 50° C. overnight and was then concentrated under reducepressure. The residue was partitioned between water and EtOAc. Theorganic extracts were washed with water, brine, dried over MgSO₄ andconcentrated to dryness to give(2-chloro-6-morpholino-9H-purin-8-yl)methanol (2.6 g, 100% yield). LC/MS(ESI+): m/z 271 (M+H)

Step 4: 2-chloro-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine

A mixture of (2-chloro-6-morpholino-9H-purin-8-yl)methanol (1 g, 0.004mol), 1,2-dibromoethane (0.64 mL, 0.0074 mol) and cesium carbonate (3.6g, 0.011 mol) in DMF (14 mL) was heated at 90° C. for 12 hours. Thereaction mixture was filtered and partitioned between water and EtOAc.The organic extracts were washed with water, brine and dried over MgSO₄and concentrated. The crude product was purified by isco with 0-50%EtOAc/hexane to give2-chloro-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine (0.7 g,60%). LC/MS (ESI+): m/z 297 (M+H)

Step 5: 2-Chloro-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(160 mg, 0.00056 mol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (180mg, 0.00083 mol) were reacted under microwave Suzuki palladiumconditions to give 113 (40 mg, 15% yield) along with some side product,(2-(2-aminopyrimidin-5-yl)-6-morpholino-9-vinyl-9H-purin-8-yl)methanol(7 mg). LC/MS (ESI+): m/z 355 (M+H). ¹H NMR (400 MHz, DMSO) δ 9.10 (s,2H), 7.00 (s, 2H), 4.91 (s, 2H), 4.34-4.07 (m, 8H), 3.90-3.63 (m, 4H).¹H NMR (400 MHz, DMSO) δ 9.10 (s, 2H), 7.35 (dd, J=15.9, 9.5 Hz, 1H),7.05 (s, 2H), 6.47 (d, J=15.9 Hz, 1H), 5.73 (t, J=5.6 Hz, 1H), 5.28 (d,J=9.4 Hz, 1H), 4.71 (d, J=5.6 Hz, 2H), 4.27 (s, 4H), 3.88-3.63 (m, 4H)

Example 1145-(4-morpholino-8,9-dihydrospiro[[1,4]oxazino[3,4-e]purine-6,3′-oxetane]-2-yl)pyrimidin-2-amine114 Step 1:3-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)oxetan-3-ol

To a mixture of4-(2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine (2.9g, 9 mmol) in THF (50 mL) at −78° C. was added 2.5 M of n-BuLi (9 mL, 22mmol) dropwise. The reaction was stirred at −78° C. for 30 minutes andthen 3-oxetanone (1.3 mL, 18 mmol) was added, continue stirred for 2hours at −78° C. The reaction was quenched with water and extracted withEtOAc. The organic extracts were washed with water, brine, dried overMgSO₄ and concentrated. The crude material was purified by Iscochromatography with 0-100% EtOAc/hexane to give pure3-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)oxetan-3-olas a white solid (2.5 g, 70% yield). LC/MS (ESI+): m/z 397 (M+H)

Step 2: 3-(2-chloro-6-morpholino-9H-purin-8-yl)oxetan-3-ol

3-(2-Chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)oxetan-3-ol(1.8 g, 0.0045 mol) in MeOH (50 mL) was treated with cat. amount ofp-toluenesulfonic acid (78 mg, 0.00044 mol). The reaction mixture washeated to 50° C. overnight and was then concentrated under reducepressure. The residue was partitioned between water and EtOAc. Theorganic extracts were washed with water, brine, dried over MgSO₄ andconcentrated to dryness to give3-(2-chloro-6-morpholino-9H-purin-8-yl)oxetan-3-ol (1.2 g, 84% yield).LC/MS (ESI+): m/z 312 (M+H)

Step 3:2-chloro-4-morpholino-8,9-dihydrospiro[[1,4]oxazino[3,4-e]purine-6,3′-oxetane]

A mixture of 3-(2-chloro-6-morpholino-9H-purin-8-yl)oxetan-3-ol (356 mg,0.0011 mol), 1,2-dibromoethane (0.21 mL, 0.0024 mol) and cesiumcarbonate (1.13 g, 0.0034 mol) in DMF (4 mL) was heated at 90° C. for 12hours. The reaction mixture was filtered and partitioned between waterand EtOAc. The organic extracts were washed with water, brine and driedover MgSO₄ and concentrated. The crude product was purified by isco with0-80% EtOAc/hexane to give2-chloro-4-morpholino-8,9-dihydrospiro[[1,4]oxazino[3,4-e]purine-6,3′-oxetane](0.34 g, 85%). LC/MS (ESI+): m/z 339 (M+H)

Step 4:2-Chloro-4-morpholino-8,9-dihydrospiro[[1,4]oxazino[3,4-e]purine-6,3′-oxetane](200 mg, 0.0006 mol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (210mg, 0.00095 mol) were reacted under microwave Suzuki palladiumconditions to give 114 (0.123 mg, 60% yield). LC/MS (ESI+): m/z 397(M+H). ¹H NMR (400 MHz, DMSO) δ 9.10 (s, 2H), 7.02 (s, 2H), 4.97 (d,J=7.1 Hz, 2H), 4.72 (d, J=7.1 Hz, 2H), 4.29 (s, 4H), 4.16 (s, 4H),3.90-3.62 (m, 4H)

Example 1155-(7,7-dimethyl-4-morpholino-8,9-dihydro-7h-[1,3]oxazino[2,3-e]purin-2-yl)pyrimidin-2-amine115 Step 1: ethyl 3-(2-chloro-6-morpholino-9H-purin-9-yl)propanoate

A solution of 4-(2-chloro-9H-purin-6-yl)morpholine (3.0 g, 13 mmol) inDMF (39 mL) was treated with cesium carbonate (8.2 g, 25 mmol) andstirred at ambient temperature for 10 minutes. 3-bromopropanoic acid,ethyl ester (6.8 g, 38 mmol) was introduced and the reaction mixture washeated at 50° C. for 2 hr. Partial conversion prompted the addition ofcesium carbonate (8.2 g, 25 mmol) and 3-bromopropanoic acid, ethyl ester(6.8 g, 38 mmol), and the reaction mixture was heated at 70° C. Thereaction mixture was cooled to ambient temperature and diluted withbrine and DCM and the layers separated. The aqueous phase was extractedinto DCM (3×), dried over sodium sulfate, filtered, and absorbed ontocelite for purification by flash chromatography to afford ethyl3-(2-chloro-6-morpholino-9H-purin-9-yl)propanoate as a white solid (3.5g, 83%). LC/MS (ESI+): m/z 340 (M+H)

Step 2: 4-(2-chloro-6-morpholino-9H-purin-9-yl)-2-methylbutan-2-ol

A solution of ethyl 3-(2-chloro-6-morpholino-9H-purin-9-yl)propanoate(500 mg, 1.5 mmol) in THF (30 mL) at 0° C. was treated drop-wise with asolution of 3.0M methylmagnesium chloride in THF (2.0 mL). After 90minutes at 0° C., the reaction mixture was treated with a saturatedsolution of NH₄Cl and diluted with brine and DCM and the layersseparated. The aqueous phase was extracted into DCM (3×), dried oversodium sulfate, filtered, and absorbed onto celite for purification byflash chromatography to afford4-(2-chloro-6-morpholino-9H-purin-9-yl)-2-methylbutan-2-ol as a whitefoam (452 mg, 94%). LC/MS (ESI+): m/z 326 (M+H)

Step 3:4-(2-chloro-8-iodo-6-morpholino-9H-purin-9-yl)-2-methylbutan-2-ol

A solution of 4-(2-chloro-6-morpholino-9H-purin-9-yl)-2-methylbutan-2-ol(360 mg, 1.1 mmol) and N, N, N′, N′-tetramethylethylenediamine (0.25 mL,1.7 mmol) in THF (8.1 mL) was cooled to −42° C. and treated with asolution of 2.5M n-butyllithium in hexane (BuLi, 2.0 mL, 5.0 mmol)dropwise over 5 minutes. After 30 minutes at −42° C.,1-chloro-2-iodoethane (0.51 mL, 5.4 mmol) was added and the reactionmixture was slowly warmed to 0° C. over 1 hr. The mixture was quenchedwith a saturated solution of NH₄Cl, and diluted with ethyl acetate. Theaqueous layer was extracted into ethyl acetate (3×), and the combinedorganics were dried over sodium sulfate, filtered, and absorbed ontocelite for purification by flash chromatography to afford4-(2-chloro-8-iodo-6-morpholino-9H-purin-9-yl)-2-methylbutan-2-ol as awhite foam (390 mg, 78%). LC/MS (ESI+): m/z 452 (M+H)

Step 4:2-chloro-7,7-dimethyl-4-morpholino-8,9-dihydro-7H-[1,3]oxazino[2,3-e]purine

Copper (1) iodide (3.2 mg, 17 umol), picolinic acid (4.1 mg, 33 umol),and potassium phosphate (140 mg, 0.67 mmol) were combined in anoven-dried round bottom flask and evacuated/recycled with N₂ (3×).Subsequently, a solution of4-(2-chloro-8-iodo-6-morpholino-9H-purin-9-yl)-2-methylbutan-2-ol (150mg, 0.33 mmol) dissolved in DMSO (2.4 mL) was introduced via syringe.The reaction mixture was heated at 80° C. for 20 hr. Partial conversionprompted the addition of copper (1) iodide (3.2 mg, 17 umol), picolinicacid (4.1 mg, 33 umol), and potassium phosphate (140 mg, 0.67 mmol) andthe reaction mixture continued to stir at 80° C. for 20 hr. The reactionmixture was cooled to ambient temperature and diluted with water andethyl acetate and the layers separated. The aqueous phase was extractedinto EtOAc (3×), dried over sodium sulfate, filtered, and absorbed ontocelite for purification by flash chromatography to afford2-chloro-7,7-dimethyl-4-morpholino-8,9-dihydro-7H-[1,3]oxazino[2,3-e]purineas a white solid (61 mg, 56%). LC/MS (ESI+): m/z 324 (M+H)

Step 5:2-Chloro-7,7-dimethyl-4-morpholino-8,9-dihydro-7H-[1,3]oxazino[2,3-e]purine(61 mg, 0.19 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (54 mg,0.24 mmol), and tetrakis(triphenylphosphine)-palladium(0) (11 mg, 9.4umol, 5.0 mol %) suspended in MeCN (0.46 mL) and 1.0M Na₂CO_(3(aq))(0.36 mL) was heated under microwave irradiation at 140° C. (for 15minutes. The cooled reaction mixture was concentrated to dryness invacuo. The resultant residue was purified via HPLC to afford 115 as awhite solid (27 mg, 37%). ¹H NMR (400 MHz, DMSO) δ 9.06 (s, 2H), 6.95(s, 2H), 4.18-4.06 (m, 6H), 3.75-3.65 (m, 4H), 2.15 (t, J=6.2 Hz, 2H),1.46 (s, 6H). LCMS: R_(T)=2.75 min, M+H⁺=383.1

Example 1165-(4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyridin-2-amine116

2-Chloro-4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine,from Examples 139 and 140 (100 mg, 0.0003 mol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) romoace-2-amine (120 mg,0.00055 mol) were reacted under microwave Suzuki palladium conditions togive 116 (56 mg, 56% yield). LC/MS (ESI+): m/z 422 (M+H). ¹H NMR (400MHz, DMSO) δ 8.94 (d, J=2.0 Hz, 1H), 8.28 (dd, J=8.7, 2.2 Hz, 1H), 6.50(d, J=8.7 Hz, 1H), 6.32 (s, 2H), 5.87 (q, J=6.9 Hz, 1H), 4.49-4.10 (m,8H), 3.75 (t, J=4.6 Hz, 4H)

Example 1175-(6,6-(hexadeuterio)dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine117 Step 1:2-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)-1,3-hexadeuterio-propan-2-ol

To a mixture of4-(2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine (5 g,0.02 mol) in THF (100 mL) at −78° C. was added 2.5 M of n-BuLi (12 mL,0.031 mol) dropwise. The reaction was stirred at −78° C. for 30 minutesand then acetone-d6 (2.5 mL, 0.034 mol) was added, continue stirred for2 hours at −78° C. The reaction was quenched with water and extractedwith EtOAc. The organic extracts were washed with water, brine, driedover MgSO₄ and concentrated. The crude material was purified by Iscochromatography with 0-100% EtOAc/hexane to give pure2-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)-1,3-hexadeuterio-propan-2-olas a white solid (5.7 g, 95% yield). LC/MS (ESI+): m/z 389 (M+H)

Step 2:2-(2-chloro-6-morpholino-9H-purin-8-yl)-1,3-hexadeuterio-propan-2-ol

2-(2-Chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)-1,3-hexadeuterio-propan-2-ol(5.7 g, 0.015 mol) in MeOH (59 mL) was treated with a catalytic amountof p-toluenesulfonic acid (253 mg, 0.00147 mol). The reaction mixturewas heated to 50° C. overnight and was then concentrated under reducepressure. The residue was partitioned between water and EtOAc. Theorganic extracts were washed with water, brine, dried over MgSO₄ andconcentrated to dryness to give2-(2-chloro-6-morpholino-9H-purin-8-yl)-1,3-hexadeuterio-propan-2-ol(4.5 g, 100% yield). LC/MS (ESI+): m/z 304 (M+H)

Step 3:2-chloro-6,6-(hexadeuterio)dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine

A mixture of2-(2-chloro-6-morpholino-9H-purin-8-yl)-1,3-hexadeuterio-propan-2-ol (2g, 0.006 mol), 1,2-dibromoethane (1.13 mL, 0.013 mol) and cesiumcarbonate (6.4 g, 0.02 mol) in DMF (4 mL) was heated at 90° C. for 12hours. The reaction mixture was filtered and partitioned between waterand EtOAc. The organic extracts were washed with water, brine and driedover MgSO₄ and concentrated. The crude product was purified by isco with0-80% EtOAc/hexane to give the pure2-chloro-6,6-(hexadeuterio)dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(1.64 g, 82%). LC/MS (ESI+): m/z 331 (M+H)

Step 4:2-Chloro-6,6-(hexadeuterio)dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(1.6 g, 0.0048 mol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (1.6 g,0.00073 mol) were reacted under Suzuki palladium conditions to give 117(600 mg, 32% yield). LC/MS (ESI+): m/z 389 (M+H). ¹H NMR (400 MHz, DMSO)δ 9.09 (s, 1H), 7.00 (s, 1H), 4.23 (s, 2H), 4.10 (t, J=13.7 Hz, 2H),3.85-3.67 (m, 2H)

Example 118(S)-5-(6-ethyl-6-methyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine118 Step 1:2-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)butan-2-ol

A solution of4-(2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine (5.0g, 15 mmol) in THF (100 mL) was cooled to −42° C. and treated with asolution of 2.5M n-Butyllithium (n-BuLi) in hexane (12.35 mL, 31 mmol)dropwise over 5 minutes. After 15 min at −42° C., 2-butanone (3.1 mL, 34mmol) was added and the reaction mixture was slowly warmed to 0° C. over2 hr. The mixture was quenched with water, and diluted with ethylacetate. The aqueous layer was extracted into ethyl acetate (3×), andthe combined organics were dried over sodium sulfate, filtered, andconcentrated in vacuo to provide2-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)butan-2-olas a yellow-orange foam (quant). LC/MS (ESI+): m/z 396 (M+H)

Step 2: 2-(2-chloro-6-morpholino-9H-purin-8-yl)butan-2-ol

A suspension of2-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)butan-2-ol(3.87 g, 9.8 mmol) in methanol (110 mL) was treated withp-toluenesulfonic acid (170 mg, 0.98 mmol) and heated overnight at 50°C. The solvent was removed in vacuo to afford2-(2-chloro-6-morpholino-9H-purin-8-yl)butan-2-ol as a white solid,which was used in the next step without any further purification (3.0 g,quant). LC/MS (ESI+): m/z 312 (M+H)

Step 3:2-chloro-6-ethyl-6-methyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine

2-(2-chloro-6-morpholino-9H-purin-8-yl)butan-2-ol (1.0 g, 3.3 mmol) wasdissolved in DMF (13 mL) and treated with 1,2-dibromoethane (0.57 mL,6.6 mmol) and cesium carbonate (3.2 g, 9.9 mmol). The reaction mixturewas heated at 90° C. for 2 hr and cooled to ambient temperature. Themixture was diluted with water and DCM and the layers separated. Theaqueous phase was extracted into DCM (3×), dried over sodium sulfate,filtered, and absorbed onto celite for purification by flashchromatography to afford2-chloro-6-ethyl-6-methyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purineas a white solid (730 mg, 66%). ¹H NMR (400 MHz, DMSO) δ 4.22 (m, 4H),4.01 (m, 2H), 3.78-3.63 (m, 4H), 2.01 (s, 2H), 1.87-1.73 (m, 2H), 1.49(s, 3H), 0.77 (t, J=7.4 Hz, 3H). LC/MS (ESI+): m/z 338 (M+H)

Step 4:2-chloro-6-ethyl-6-methyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(730 mg, 2.2 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (620mg, 2.8 mmol), and tetrakis(triphenylphosphine)palladium(0) (120 mg,0.11 mmol, 5.0 mol %) suspended in MeCN (5.2 mL) and 1.0M Na₂CO_(3(aq))(4.1 mL) was heated under microwave irradiation at 140° C. for 15minutes. The cooled reaction mixture was concentrated to dryness invacuo. The resultant residue was purified by SFC (conditions A) over 30min, 35 mL/min] to separate the two enantiomers 118 and 120 to afford118 as a white solid (120 mg and 116 mg, 30%). ¹H NMR (400 MHz, DMSO) δ9.09 (s, 2H), 7.00 (s, 2H), 4.29-4.01 (m, 8H), 3.80-3.68 (m, 4H), 2.00(dq, J=14.5, 7.3 Hz, 1H), 1.84 (dt, J=14.4, 7.2 Hz, 1H), 1.52 (s, 3H),0.82 (t, J=7.3 Hz, 3H). LCMS: R_(T)=9.49 min, M+H⁺=397.1. Enantiomers118 and 120 were analyzed and separated by chiral LCMS, rt=1.20 min and1.65 min with mobile phase A=C02, mobile phase B=methanol, isocratic 25%B, 5 ml/min flow rate, 40° C., ChiralCel OJ (4.6×50 mm, 3 micronparticle, 230 nm UV detection, Berger Analytical SFC/MS

Example 1195-(6,6,9-trimethyl-4-morpholino-6h-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine119 Step 1: methyl2-(2-chloro-8-(2-hydroxypropan-2-yl)-6-morpholino-9H-purin-9-yl)propanoate

2-(2-chloro-6-morpholino-9H-purin-8-yl)propan-2-ol (4.6 g, 15 mmol) wasdissolved in DMF (16 mL), treated with cesium carbonate (10 g, 31 mmol)and methyl 2-bromopropanoate (7.6 g, 46 mmol), and heated at 50° C. for3 hr. Partial conversion prompted addition of cesium carbonate (10 g, 31mmol) and methyl 2-bromopropanoate (7.6 g, 46 mmol) and the reactionmixture was heated at 50° C. for 20 hr. The reaction mixture was cooledto ambient temperature and diluted with water and ethyl acetate and thelayers separated. The aqueous phase was extracted into ethyl acetate(3×), dried over sodium sulfate, filtered, and absorbed onto celite forpurification by flash chromatography to afford methyl2-(2-chloro-8-(2-hydroxypropan-2-yl)-6-morpholino-9H-purin-9-yl)propanoateas a yellow foam (1.6 g, 26%). LC/MS (ESI+): m/z 384 (M+H)

Step 2:2-(2-chloro-8-(2-hydroxypropan-2-yl)-6-morpholino-9H-purin-9-yl)propanal

Methyl2-(2-chloro-8-(2-hydroxypropan-2-yl)-6-morpholino-9H-purin-9-yl)propanoate(1.6 g, 4.1 mmol) was dissolved in THF (30 mL) and cooled to −78° C. Thereaction mixture was treated with a 1.0M solution of lithiumtetrahydroaluminate in THF (8.6 mL) and stirred at −78° C. for 1 hr. Asaturated solution of NH4Cl was added and the mixture was diluted withDCM. The reaction mixture was treated with a saturated solution ofRochelle's salt and stirred on high at ambient temperature for 1 hr. Thelayers were separated and the aqueous phase was extracted into DCM/MeOH(3×), dried over sodium sulfate, filtered, and absorbed onto celite forpurification by flash chromatography to afford2-(2-chloro-8-(2-hydroxypropan-2-yl)-6-morpholino-9H-purin-9-yl)propanalas a white foam (900 mg, 62%). LC/MS (ESI+): m/z 384 (M+H)

Step 3:2-chloro-6,6,9-trimethyl-4-morpholino-6H-[1,4]oxazino[3,4-e]purine

A solution of2-(2-chloro-8-(2-hydroxypropan-2-yl)-6-morpholino-9H-purin-9-yl)propanal(900 mg, 2.5 mmol) in toluene (8.1 mL), was treated with trifluoroaceticacid (0.58 mL, 7.6 mmol) and heated at 110° C. for 4 hr. Partialconversion prompted addition of trifluoroacetic acid (1.0 mL) and themixture was allowed to stir overnight at 110° C. The reaction mixturewas cooled to ambient temperature and concentrated to dryness in vacuo.The resultant residue was re-dissolved in DCM and absorbed onto celitefor purification by flash chromatography to afford2-chloro-6,6,9-trimethyl-4-morpholino-6H-[1,4]oxazino[3,4-e]purine as asolid (190 mg, 22%). LC/MS (ESI+): m/z 336/338 (M+H)

Step 4:2-chloro-6,6,9-trimethyl-4-morpholino-6H-[1,4]oxazino[3,4-e]purine (190mg, 0.55 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (160mg, 0.71 mmol), and tetrakis(triphenylphosphine)-palladium(0) (32 mg, 27umol, 5.0 mol %) suspended in MeCN (1.3 mL) and 1.0M Na₂CO_(3(aq)) (1.0mL) was heated under microwave irradiation at 140° C. for 15 minutes.The cooled reaction mixture was concentrated to dryness in vacuo. Theresultant residue was purified via HPLC to afford 119 as a white solid(120 mg, 54%). LC/MS (ESI+): m/z 395 (M+H). ¹H NMR (400 MHz, DMSO) δ9.06 (s, 2H), 7.03 (s, 2H), 6.34 (s, 1H), 4.24 (s, 4H), 3.80-3.70 (m,4H), 2.52 (s, 3H), 1.62 (s, 6H). LCMS: R_(T)=4.57 min, M+H⁺=395.2

Example 120(R)-5-(6-ethyl-6-methyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine120

Following the procedures of Example 118, the R-enantiomer 120 wasisolated.

Example 1215-(1-morpholin-4-yl-5,6,8a,9-tetrahydro-8h-7,10-dioxa-2,4,4b-triaza-phenanthren-3-yl)-pyrimidin-2-ylamine121 Step 1:[4-(2,6-Dichloro-5-methoxy-pyrimidin-4-yl)-morpholin-3-yl]-methanol

A mixture of 2,4,6-trichloro-5-methoxy-pyrimidine (1 g, 4.68 mmol),morpholin-3-yl-methanol hydrochloride (0.86 g, 5.6 mmol) andtriethylamine (0.9 mL, 6.5 mmol) in IMS (30 mL) was stirred at RT for 3h, then concentrated in vacuo. The resulting residue was purified bycolumn chromatography (SiO₂, 0 to 50% ethyl acetate in cyclohexane)affording[4-(2,6-Dichloro-5-methoxy-pyrimidin-4-yl)-morpholin-3-yl]-methanol (614mg, 45%). LCMS (method A): R_(T)=2.63 min, [M+H]⁺=294/296.

Step 2:1,3-Dichloro-5,6,8a,9-tetrahydro-8H-7,10-dioxa-2,4,4b-triaza-phenanthrene

A mixture of[4-(2,6-dichloro-5-methoxy-pyrimidin-4-yl)-morpholin-3-yl]-methanol (614mg, 2.09 mmol) and lithium chloride (246 mg, 5.80 mmol) in anhydrous DMF(5 mL) was heated at 160° C. for 10 mins in a microwave reactor, thenconcentrated in vacuo to give2,4-dichloro-6-(3-hydroxymethyl-morpholin-4-yl)-pyrimidin-5-ol. DIAD(452 μL, 2.3 mmol) was added to a solution of2,4-dichloro-6-(3-hydroxymethyl-morpholin-4-yl)-pyrimidin-5-ol (2 mmol)and triphenyl phosphine (603 mg, 2.3 mmol) in 1,4-dioxane (5 mL) and themixture stirred at RT for 1 h, then concentrated in vacuo. The resultingresidue was purified by column chromatography (SiO₂, 0 to 50% ethylacetate in cyclohexane) affording1,3-Dichloro-5,6,8a,9-tetrahydro-8H-7,10-dioxa-2,4,4b-triaza-phenanthrene(200 mg, 37%). LCMS (method A): R_(T)=2.91 min, [M+H]⁺=262/264.

Step 3:3-Chloro-1-morpholin-4-yl-5,6,8a,9-tetrahydro-8H-7,10-dioxa-2,4,4b-triaza-phenanthrene

A mixture of1,3-dichloro-5,6,8a,9-tetrahydro-8H-7,10-dioxa-2,4,4b-triaza-phenanthrene(100 mg, 0.38 mmol), morpholine (80 μL, 0.92 mmol) and triethylamine (70μL, 0.50 mmol) in IMS (5 mL) was heated at 140° C. for 25 mins in amicrowave reactor, then concentrated in vacuo. The resulting residue waspurified by column chromatography (SiO₂, 0 to 10 to 15 to 25% ethylacetate in pentane) affording3-Chloro-1-morpholin-4-yl-5,6,8a,9-tetrahydro-8H-7,10-dioxa-2,4,4b-triaza-phenanthrene(45 mg, 38%). LCMS (method A): R_(T)=2.92 min, [M+H]⁺=313. ¹H NMR (400MHz, CDCl₃): δ 4.37 (1H, dd, J=13.2, 3.0 Hz), 4.19 (1H, dd, J=10.8, 3.3Hz), 3.99 (1H, dd, J=11.1, 3.9 Hz), 3.88 (1H, dd, J=10.8, 3.2 Hz), 3.80(dd, J=11.1, 8.4 Hz), 3.75 (4H, m), 3.66-3.53 (6H, m), 3.24 (1H, t,J=11.1 Hz), 3.00 (1H, m).

Step 4: A mixture of3-chloro-1-morpholin-4-yl-5,6,8a,9-tetrahydro-8H-7,10-dioxa-2,4,4b-triaza-phenanthrene(45 mg, 0.144 mmol),5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrimidin-2-ylamine (60mg, 0.271 mmol), PdCl₂(PPh₃)₂(10 mg, 0.014 mmol) and sodium carbonate(460 μL, 0.46 mmol, 1M aqueous solution) in acetonitrile (2 mL) wasdegassed and heated at 120° C. for 30 mins in a microwave reactor. Thereaction mixture was loaded onto an Isolute® SCX-2 cartridge which waswashed with methanol and the product eluted with 2M ammonia in methanol.The basic fractions were combined and concentrated in vacuo. Theresulting residue was purified by reverse phase HPLC (Phenomenex Gemini5 μm C18, 0.1% HCO₂H in water on a gradient acetonitrile 5-98%)affording 121 as a white solid (3 mg, 6%). LCMS (method B): R_(T)=3.08min, [M+H]⁺=372. ¹H NMR (400 MHz, CDCl₃): δ 9.11 (2H, s), 5.18 (2H,broad s), 4.55 (1H, dd, J=13.5, 2.4 Hz), 4.22 (1H, dd, J=10.8, 3.1 Hz),4.05 (1H, dd, J=11.5, 3.5 Hz), 3.88 (2H, m), 3.81 (4H, m), 3.69-3.56(6H, m), 3.29 (1H, t, J=11.5 Hz), 3.02 (1H, m).

Example 1225-((S)-6-Morpholin-4-yl-2,3,3a,4-tetrahydro-1H-5-oxa-7,9,9b-triaza-cyclopenta[a]naphthalen-8-yl)-pyrimidin-2-ylamine122 Step 1:[(S)-1-(2,6-Dichloro-5-methoxy-pyrimidin-4-yl)-pyrrolidin-2-yl]-methanol

A mixture of 2,4,6-trichloro-5-methoxy-pyrimidine (1.2 g, 5.62 mmol),(S)-1-pyrrolidin-2-yl-methanol (1.1 mL, 11.3 mmol) and triethylamine(1.08 mL, 7.75 mmol) in IMS (36 mL) was stirred at RT for 20 mins, thenconcentrated in vacuo. The resulting residue was purified by columnchromatography (SiO₂, 0 to 50% ethyl acetate in cyclohexane) affording[(S)-1-(2,6-Dichloro-5-methoxy-pyrimidin-4-yl)-pyrrolidin-2-yl]-methanol(1.08 mg, 70%). LCMS (method A): R_(T)=2.98 min, [M+H]⁺=278/280.

Step 2:(S)-6,8-Dichloro-2,3,3a,4-tetrahydro-1H-5-oxa-7,9,9b-triaza-cyclopenta[a]naphthalene

A mixture of[(S)-1-(2,6-dichloro-5-methoxy-pyrimidin-4-yl)-pyrrolidin-2-yl]-methanol(900 mg, 3.24 mmol) and lithium chloride (360 mg, 8.48 mmol) inanhydrous DMF (10 mL) was heated at 160° C. for 10 mins in a microwavereactor, then concentrated in vacuo to give2,4-dichloro-6-((S)-2-hydroxymethyl-pyrrolidin-1-yl)-pyrimidin-5-ol.DIAD (700 μL, 3.56 mmol) was added to a solution of2,4-dichloro-6-((S)-2-hydroxymethyl-pyrrolidin-1-yl)-pyrimidin-5-ol (3mmol) and triphenyl phosphine (900 mg, 3.43 mmol) in 1,4-dioxane (10 mL)and the mixture stirred at RT for 1 h, then concentrated in vacuo. Theresulting residue was purified by column chromatography (SiO₂, 0 to 20%ethyl acetate in cyclohexane) affording(S)-6,8-dichloro-2,3,3a,4-tetrahydro-1H-5-oxa-7,9,9b-triaza-cyclopenta[a]naphthalene.LCMS (method A): R_(T)=2.98 min, [M+H]⁺=246/248.

Step 3: A mixture of(S)-6,8-dichloro-2,3,3a,4-tetrahydro-1H-5-oxa-7,9,9b-triaza-cyclopenta[a]naphthalene(290 mg, 1.18 mmol), morpholine (275 μL, 3.14 mmol) and triethylamine(242 μL, 1.74 mmol) in IMS (11 mL) was heated at 140° C. for 20 mins ina microwave reactor, then concentrated in vacuo. The resulting residuere-dissolved in acetonitrile (2 mL) and5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrimidin-2-ylamine(500 mg, 2.26 mmol), PdCl₂(PPh₃)₂(88 mg, 0.125 mmol) and sodiumcarbonate (4 mL, 4.0 mmol, 1M aqueous solution) were added. The reactionmixture was degassed and heated at 120° C. for 30 mins in a microwavereactor. The reaction mixture was loaded onto an Isolute® an SCX-2cartridge which was washed with methanol and the product eluted with 2Mammonia in methanol. The basic fractions were combined and concentratedin vacuo. The resulting residue was purified by reverse phase HPLC(Phenomenex Gemini 5 μm C18, 0.1% HCO₂H in water gradient acetonitrile5-60%) affording 122 as a white solid (30 mg, 8%). LCMS (method B):R_(T)=3.34 min, [M+H]⁺=356. ¹H NMR (400 MHz, CDCl₃): δ 9.15 (2H, s),5.14 (2H, broad s), 4.48 (1H, dd, J=10.4, 3.5 Hz), 3.87-3.68 (10H, m),3.62 (1H, m), 3.31 (1H, t, J=10.1 Hz), 2.21-1.94 (3H, m), 1.50 (1H, m).

Also isolated was the regioisomer,5-((S)-8-morpholin-4-yl-2,3,3a,4-tetrahydro-1H-5-oxa-7,9,9b-triaza-cyclopenta[a]romoacetyl-6-yl)-pyrimidin-2-ylamine (25 mg, 6%). LCMS (method B):R_(T)=2.47 min, [M+H]⁺=356. ¹H NMR (400 MHz, CDCl₃): δ 9.09 (2H, s),5.14 (2H, broad s), 4.49 (1H, dd, J=10.8, 3.8 Hz), 3.80-3.65 (10H, m),3.66 (1H, m), 3.34 (1H, t, J=9.7 Hz), 2.20-1.94 (3H, m), 1.48 (1H, m).

Example 1234-(6,6-dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)aniline123

To2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purinefrom Example 103 and following General Procedure A (266 mg, 0.82 mmol)in acetonitrile (2.5 mL) was added 4-aminophenylboronic acid, pinacolester (270 mg, 1.2 mmol) and 1.0 M of cesium carbonate in water (2.5mL). The reaction mixture was degassed for 5 min and recycled withnitrogen atmosphere. Subsequently,bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)palladium(II)dichloride (29 mg, 0.041 mmol) was added, and the mixture was degassedand recycled again. The reaction vial was then subjected to microwaveirradiation for 25 mins at 100° C. The vessel was cooled to roomtemperature and extracted twice with EtOAc. Dried over MgSO₄, filteredand concentrated in vacuo. Purified by rp-HPLC to provide 123 (151 mg,48% yield). MS (ESI+): m/z 381.2 (M+H⁺). ¹H NMR (400 MHz, DMSO) δ 8.08(d, J=8.5 Hz, 1H), 6.59 (d, J=8.5 Hz, 1H), 5.42 (s, 1H), 4.22 (s, 2H),4.11 (s, 2H), 3.80-3.67 (nm, 2H), 1.57 (s, 31H)

Example 1241-(4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)-3-methylurea124

To a solution of4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)aniline123 (0.44 g, 1.2 mmol) in 1,2-dichloroethane (10 mL) was addedtriethylamine (0.35 mL, 2.5 mmol) and the reaction mixture was cooled to0° C. Triphosgene (0.17 g, 0.57 mmol) was added slowly and the mixturewas subsequently warmed to 70° C. for 1 h. The reaction mixture was thencooled to room temperature for the addition of 2.0 M of methylamine inTHF (2.2 mL, 4.4 mmol) and the resulting reaction mixture was stirredfor 16 h at ambient temperature. LC-MS indicated complete conversion andas a result the reaction mixture was diluted with water and EtOAc. Thephases were separated and the aqueous layer was extracted 3× with EtOAc.The organic extracts were collected and dried over MgSO₄, filtered andconcentrated in vacuo. Purified by rp-HPLC to provide 124 (122 mg, 25%yield). MS (ESI+): m/z 438.2 (M+H⁺). ¹H NMR (400 MHz, DMSO) δ 8.68 (s,1H), 8.24 (d, J=8.7 Hz, 2H), 7.48 (d, J=8.7 Hz, 2H), 6.04 (q, J=4.4 Hz,1H), 4.25 (s, 4H), 4.14 (d, J=3.4 Hz, 4H), 3.82-3.67 (m, 4H), 2.66 (d,J=4.6 Hz, 3H), 1.58 (s, 6H)

Example 1256,6-dimethyl-4-morpholino-2-(1H-pyrazol-4-yl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine125

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.31mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 125. 1H NMR (400 MHz, DMSO) δ12.98 (s, 1H), 8.23 (s, 1H), 8.01 (s, 1H), 4.22 (s, 4H), 4.10 (s, 4H),3.80-3.67 (m, 4H), 1.57 (s, 6H). LCMS: R_(T)=3.67 min, M+H⁺=356

Example 1264-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyridin-2-amine126

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)romoace-2-amine (0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 126. 1H NMR (400 MHz, DMSO) δ 7.99(d, J=5.3 Hz, 1H), 7.41 (s, 1H), 7.38 (d, J=5.3 Hz, 1H), 5.97 (s, 2H),4.27 (s, 4H), 4.15 (d, J=3.5 Hz, 4H), 3.82-3.71 (m, 4H), 1.59 (s, 6H).LCMS: R_(T)=3.64 min, M+H⁺=382

Example 1276,6-dimethyl-2-(1-methyl-1H-pyrazol-4-yl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine127

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 127. 1H NMR (400 MHz, DMSO) δ 8.23(s, 1H), 7.93 (s, 1H), 4.21 (s, 4H), 4.10 (s, 41H), 3.88 (s, 3H), 3.73(dd, J=12.3, 7.7 Hz, 4H), 1.57 (s, 6H). LCMS: R_(T)=3.94 min, M+H⁺=370

Example 1283-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenol128

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 128. 1H NMR (400 MHz, DMSO) δ 9.41(s, 1H), 7.82 (dd, J=4.5, 2.5 Hz, 2H), 7.28-7.20 (m, 1H), 6.82 (dd,J=7.7, 1.8 Hz, 1H), 4.26 (s, 4H), 4.19-4.08 (m, 4H), 3.82-3.71 (m, 4H),1.59 (s, 6H). LCMS: R_(T)=4.46 min, M+H⁺=382

Example 1292-(1H-indazol-5-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine129

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (0.31mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 129. 1H NMR (400 MHz, DMSO) δ13.11 (s, 1H), 8.81 (s, 1H), 8.48-8.43 (m, 1H), 8.19 (s, 1H), 7.58 (d,J=8.8 Hz, 1H), 4.29 (s, 4H), 4.17 (dd, J=16.1, 5.1 Hz, 4H), 3.82-3.74(m, 4H), 1.60 (s, 6H). LCMS: R_(T)=4.47 min, M+H⁺=406

Example 130 6,6-dimethyl-2-(2-(4-methylpiperazin-1-yl)romoace-4-yl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine 130

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)romoace-2-yl)piperazine (0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 130.111 NMR (400 MHz, DMSO) δ 8.21(d, J=4.7 Hz, 1H), 7.68 (s, 1H), 7.56 (d, J=4.7 Hz, 1H), 4.27 (s, 4H),4.16 (d, J=26.1 Hz, 4H), 3.77 (s, 4H), 3.55 (s, 4H), 2.46 (s, 4H), 2.25(s, 3H), 1.59 (s, 6H). LCMS: R_(T)=3.40 min, M+H⁺=465

Example 131N-(2-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)methanesulfonamide131

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),N-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanesulfonamide(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 131. 1H NMR (400 MHz, DMSO) δ12.87 (s, 1H), 8.58 (d, J=8.0 Hz, 1H), 7.60 (d, J=8.1 Hz, 1H), 7.48 (t,J=7.5 Hz, 1H), 7.22 (t, J=7.4 Hz, 1H), 4.26 (s, 4H), 4.16 (s, 4H), 3.78(s, 4H), 3.07 (s, 3H), 1.62 (d, J=11.5 Hz, 6H). LCMS: R_(T)=5.36 min,M+H⁺=459

Example 1326,6-dimethyl-4-morpholino-2-(6-morpholinopyridin-3-yl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine132

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol), 4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)romoace-2-yl)morpholine (0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 132. 1H NMR (400 MHz, DMSO) δ 9.11(s, 1H), 8.42 (d, J=8.7 Hz, 1H), 6.89 (d, J=8.9 Hz, 1H), 4.24 (s, 4H),4.14 (s, 4H), 3.74 (d, J=14.7 Hz, 8H), 3.55 (d, J=3.9 Hz, 41H), 1.60 (d,J=15.6 Hz, 6H). LCMS: R_(T)=3.79 min, M+H⁺=452

Example 1332-(1-benzyl-1H-pyrazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine133

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),1-benzyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 133. 1H NMR (400 MHz, DMSO) δ 8.37(s, 1H), 7.99 (s, 1H), 7.35 (d, J=7.6 Hz, 2H), 7.29 (d, J=6.9 Hz, 3H),5.37 (s, 2H), 4.21 (s, 4H), 4.09 (s, 4H), 3.74 (s, 4H), 1.58 (d, J=9.4Hz, 6H). LCMS: R_(T)=4.82 min, M+H⁺=446

Example 1342-(2-isopropoxypyridin-3-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine134

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),2-isopropoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 134. 1H NMR (400 MHz, DMSO) δ 8.20(s, 1H), 8.03 (d, J=7.2 Hz, 1H), 7.07-6.99 (m, 1H), 5.43-5.31 (m, 1H),4.22 (s, 4H), 4.11 (s, 4H), 3.73 (s, 4H), 1.59 (s, 6H), 1.26 (d, J=6.1Hz, 6H). LCMS: R_(T)=4.56 min, M+H⁺=425

Example 135N-(2-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)acetamide135

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),N-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetamide (0.31mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 135. 1H NMR (400 MHz, DMSO) δ12.45 (s, 1H), 8.53 (d, J=8.2 Hz, 1H), 8.44 (d, J=8.2 Hz, 1H), 7.40 (t,J=7.5 Hz, 1H), 7.15 (t, J=7.5 Hz, 1H), 4.23 (t, J==18.4 Hz, 41), 4.16(s, 2H), 3.78 (s, 4-1), 2.21 (s, 3H), 1.62 (d, J=10.6 Hz, 6H). LCMS:R_(T)=5.16 min, M+H⁺=423

Example 1362-(3,5-dimethyl-1H-pyrazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine136

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 136. LCMS: R_(T)=3.81 min,M+H⁺=384

Example 1375-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)romoace-2-ol 137

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)romoace-2-ol (0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 137. 1H NMR (400 MHz, DMSO) δ11.82 (s, 1H), 8.34 (d, J=9.7 Hz, 1H), 8.27 (s, 1H), 6.41 (d, J=9.6 Hz,1H), 4.22 (s, 4H), 4.12 (s, 4H), 3.75 (s, 4H), 1.57 (s, 6H). LCMS:R_(T)=3.95 min, M+H⁺=383

Example 1386-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)romoace-3-amine 138

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol), 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)romoace-3-amine (0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 138. 1H NMR (400 MHz, DMSO) δ 9.20(s, 1H), 8.91 (s, 1H), 8.22 (d, J=8.4 Hz, 1H), 8.07 (d, J=3.8 Hz, 1H),7.25 (dd, J=8.1, 4.3 Hz, 1H), 4.25-3.97 (m, 8H), 3.74 (d, J=3.9 Hz, 4H),1.55 (s, 6H). LCMS: R_(T)=3.44 min, M+H⁺=382

Examples 139 and 140(R)-5-(4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine139 and(S)-5-(4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine140 Step 1:1-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)-2,2,2-trifluoroethanone

To a mixture of4-(2-chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-yl)morpholine (1 g,9 mmol) in THF (25 mL) at −78° C. was added Tetramethylethylenediamine(0.93 mL, 0.0062 mol) followed by 2.5 M of n-BuLi (2.5 mL, 0.0062 mol)dropwise. The reaction was stirred at −78° C. for 30 minutes and thenethyl trifluoroacetate (0.74 mL, 0.0062 mol) was added, continue stirredfor 2 hours at −78° C. The reaction was quenched with water andextracted with EtOAc. The organic extracts were washed with water,brine, dried over MgSO₄ and concentrated. The crude material waspurified by ISCO with 0-100%, EtOAc/hexane to give pure1-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)-2,2,2-trifluoroethanoneas a white solid (2.5 g, 70% yield). LC/MS (ESI+): m/z 421 (M+H)

Step 2:1-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)-2,2,2-trifluoroethanol

1-(2-Chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)-2,2,2-trifluoroethanone(1.5 g, 0.0036 mol) in MeOH (22 mL) was treated with sodiumtetrahydroborate (0.27 g, 0.0072 mol), and stirred at room temperaturefor 1 hour. The reaction was quenched with water and extracted withEtOAc. The organic extracts were washed with water, brine, dried overMgSO₄ and concentrated. The crude material was purified with ISCO with0-80% EtOAc/hexane to give the pure1-(2-chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)-2,2,2-trifluoroethanol(1.3 g, 86% yield). LC/MS (ESI+): m/z 423 (M+H)

Step 3: 1-(2-chloro-6-morpholino-9H-purin-8-yl)-2,2,2-trifluoroethanol

1-(2-Chloro-6-morpholino-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-8-yl)-2,2,2-trifluoroethanol(1.3 g, 0.0031 mol) in MeOH (12 mL) was treated with cat. amount ofp-toluenesulfonic acid (53 mg, 0.00031 mol). The reaction mixture washeated to 50° C. overnight and was then concentrated under reducepressure. The residue was partitioned between water and EtOAc. Theorganic extracts were washed with water, brine, dried over MgSO₄ andconcentrated to dryness to give1-(2-chloro-6-morpholino-9H-purin-8-yl)-2,2,2-trifluoroethanol (1 g,100% yield). LC/MS (ESI+): m/z 338 (M+H)

Step 4:2-chloro-4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine

A mixture of1-(2-chloro-6-morpholino-9H-purin-8-yl)-2,2,2-trifluoroethanol (1 g,0.003 mol), 1,2-dibromoethane (0.51 mL, 0.006 mol) and cesium carbonate(2.9 g, 0.089 mol) in DMF (18 mL) was heated at 90′C for 12 hours. Thereaction mixture was filtered and partitioned between water and EtOAc.The organic extracts were washed with water, brine and dried over MgSO₄and concentrated. The crude product was purified by isco with 0-50%EtOAc/hexane to give the pure2-chloro-4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(0.3 g, 30%). LC/MS (ESI+): m/z 364 (M+H)

Step 5:2-Chloro-4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(140 mg, 0.0004 mol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (130mg, 0.00058 mol) were reacted under microwave Suzuki palladiumconditions to give the racemic5-(4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine(36 mg, 32% yield) which was separated into the (R) enantiomer 139 andthe (S) enantiomer 140. LC/MS (ESI+): m/z 423 (M+H). ¹H NMR (400 MHz,DMSO) δ 9.11 (s, 2H), 7.05 (s, 2H), 5.88 (d, J=6.8 Hz, 1H), 4.38 (t,J=12.2 Hz, 2H), 4.35-4.09 (m, 6H), 3.76 (s, 4H)

Example 1412-(1-ethyl-1H-pyrazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine141

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),1-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 141. 1H NMR (400 MHz, DMSO) δ 8.26(s, 1H), 7.95 (s, 1H), 4.28-4.12 (m, 6H), 4.10 (s, 4H), 3.78-3.69 (m,4H), 1.58 (s, 6H), 1.40 (t, J=7.3 Hz, 3H). LCMS: R_(T)=4.13 min,M+H⁺=384

Example 1424-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)-N,N-dimethylbenzamide142

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),N,N-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (1)(1 mg, 16 umol) were reacted to give 142. 1H NMR (400 MHz, DMSO) δ 8.42(d, J=8.2 Hz, 2H), 7.49 (d, J=8.2 Hz, 2H), 4.28 (s, 4H), 4.16 (m, 4H),3.81-3.73 (m, 4H), 3.06-2.87 (m, 6H), 1.60 (s, 6H). LCMS: R_(T)=4.60min, M+H⁺=437

Example 143 tert-butyl4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl(methyl)carbamate143

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol), tert-butylmethyl(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)carbamate(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 143. LCMS: R_(T)=6.07 min,M+H⁺=495

Example 1442-(3-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)acetonitrile144

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetonitrile(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 144. 1H NMR (400 MHz, DMSO) δ 8.37(s, 1H), 8.35 (d, J=7.8 Hz, 1H), 7.50 (t, J=7.6 Hz, 1H), 7.43 (d, J=7.5Hz, 1H), 4.28 (s, 4H), 4.21-4.1 (m, 6H), 3.84-3.70 (m, 4H), 1.59 (s,6H). LCMS: R_(T)=5.11 min, M+H⁺=405 11.7

Example 1456,6-dimethyl-4-morpholino-2-(3-morpholinophenyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine145

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)morpholine(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 145. 1H NMR (400 MHz, DMSO) δ 7.96(s, 1H), 7.86 (d, J=7.7 Hz, 1H), 7.32 (t, J=7.9 Hz, 1H), 7.04 (dd,J=8.1, 2.2 Hz, 1H), 4.25 (s, 4H), 4.15 (m, 4H), 3.77 (m, 8H), 3.21-3.12(m, 4H), 1.59 (s, 6H). LCMS: R_(T)r=4.66 min, M+H⁺=451

Example 1466,6-dimethyl-4-morpholino-2-(3-(morpholinomethyl)phenyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine146

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 146. 1H NMR (400 MHz, DMSO) δ 8.30(s, 1H), 8.27 (d, J=7.3 Hz, 1H), 7.46-7.34 (m, 2H), 4.27 (s, 4H), 4.6(m, 4H), 3.83-3.72 (m, 4H), 3.58 (m, 4H), 3.55 (s, 2H), 2.38 (m, 4H),1.59 (s, 6H). LCMS: R_(T)=3.84 min, M+H⁺=465

Example 1472-(3-(benzyloxy)phenyl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine147

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),2-(3-(benzyloxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.31mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 147. 1H NMR (400 MHz, DMSO) δ8.00-7.94 (m, 2H), 7.50 (d, J=7.2 Hz, 2H), 7.44-7.37 (m, 3H), 7.37-7.30(m, 1H), 7.14-7.07 (m, 1H), 5.20 (s, 2H), 4.21 (s, 4H), 4.15 (m, 4H),3.82-3.69 (m, 4H), 1.59 (s, 6H). LCMS: R_(T)=6.33 min, M+H⁺=472

Example 1482-(1-isobutyl-1H-pyrazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine148

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),1-isobutyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(!1 mg, 16 umol) were reacted to give 148. 1H NMR (400 MHz, DMSO) δ 8.24(s, 1H), 7.95 (d, J=8.5 Hz, 1H), 4.22 (s, 4H), 4.10 (s, 4H), 3.95 (d,J=7.2 Hz, 2H), 3.78-3.69 (m, 4H), 2.15 (dp, J=13.8, 6.8 Hz, 1H), 1.57(s, 6H), 0.86 (d, J=6.7 Hz, 6H). LCMS: R_(T)=4.67 min, M+H⁺=412

Example 149 6,6-dimethyl-2-(6-(4-methylpiperazin-1-yl)romoace-3-yl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine 149

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),1-methyl-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)romoace-2-yl)piperazine (0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 149. 1H NMR (400 MHz, DMSO) δ 9.08(d, J=2.3 Hz, 1H), 8.39 (dd, J=9.0, 2.3 Hz, 1H), 6.89 (t, J=7.1 Hz, 1H),4.24 (s, 4H), 4.14 (t, J=5.2 Hz, 4H), 3.81-3.70 (m, 4H), 3.64-3.52 (m,4H), 2.44-2.36 (m, 4H), 2.23 (s, 3H), 1.58 (s, 6H). LCMS: R_(T)=3.45min, M+H⁺=465

Example 1502-(1H-indazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine150

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole 24 (0.31mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 150. 1H NMR (400 MHz, DMSO) δ13.16 (d, J=20.9 Hz, 1H), 8.91 (s, 1H), 8.20 (d, J=7.2 Hz, 1H), 7.64 (t,J=8.3 Hz, 1H), 7.45 (t, J=7.8 Hz, 1H), 4.28 (m, 6H), 4.17 (m, 2H),3.85-3.75 (m, 4H), 1.61 (s, 6H). LCMS: R_(T)=4.61 min, M+H⁺=406

Example 1514-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)benzonitrile151

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (0.31mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 151. 1H NMR (400 MHz, DMSO) δ 8.54(d, J=8.5 Hz, 2H), 7.94 (d, J=8.4 Hz, 2H), 4.28 (m, 4H), 4.16 (m, 4H),3.77 (m, 4H), 1.60 (s, 61H). LCMS: R_(T)=5.53 min, M+H⁺=391

Example 1525-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)nicotinamide152

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)nicotinamide (0.31mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 152. 1H NMR (400 MHz, DMSO) δ 9.61(dd, J=5.2, 2.0 Hz, 1H), 9.08 (t, J=2.7 Hz, 1H), 9.02 (t, J=2.1 Hz, 1H),8.30 (s, 1H), 7.65 (s, 1H), 4.30 (s, 4H), 4.18 (dt, J=9.7, 4.5 Hz, 4H),3.81-3.71 (m, 4H), 1.60 (s, 6H). LCMS: R_(T)=3.71 min, M+H⁺=410

Example 1535-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)-N-methylpicolinamide153

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),N-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamide(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 153. 1H NMR (400 MHz, DMSO) δ 9.51(s, 1H), 8.81 (d, J=6.1 Hz, 1H), 8.79-8.72 (m, 1H), 8.13 (d, J=8.2 Hz,1H), 4.28 (m, 4H), 4.17 (m, 4H), 3.78 (m, 4H), 2.86 (d, J=4.9 Hz, 3H),1.60 (s, 6H). LCMS: R_(T)=4.64 min, M+H⁺=424

Example 1542-(4-(benzyloxy)phenyl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine154

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),2-(4-(benzyloxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.31mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 154. 1H NMR (400 MHz, DMSO) δ 8.31(t, J=7.6 Hz, 2H), 7.48 (d, J=7.2 Hz, 2H), 7.41 (t, J=7.4 Hz, 2H), 7.34(t, J=7.2 Hz, 1H), 7.10 (t, J=7.4 Hz, 2H), 5.17 (s, 21H), 4.25 (s, 4H),4.14 (dd, J=6.7, 2.6 Hz, 4H), 3.81-3.71 (m, 4H), 1.58 (s, 6H). LCMS:R_(T)=6.21 min, M+H⁺=472

Example 1553-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)-N,N-dimethylaniline155

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),N,N-dimethyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 155. 1H NMR (400 MHz, DMSO) δ 7.80(s, 1H), 7.71 (d, J=7.7 Hz, 1H), 7.26 (t, J=7.9 Hz, 1H), 6.82 (dd,J=8.2, 2.5 Hz, 1H), 4.25 (s, 4H), 4.23-4.06 (m, 4H), 3.84-3.69 (m, 4H),2.96 (s, 6H), 1.59 (s, 6H). LCMS: R_(T)=3.88 min, M+H⁺=409

Example 1566,6-dimethyl-2-(4-(4-methylpiperazin-1-yl)phenyl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine156

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 156. 1H NMR (400 MHz, DMSO) δ 8.22(d, J=8.9 Hz, 2H), 6.98 (d, J=8.9 Hz, 2H), 4.24 (s, 4H), 4.13 (d, J=3.2Hz, 4H), 3.80-3.72 (m, 4H), 3.26-3.20 (m, 4H), 2.48-2.43 (m, 4H), 2.23(s, 3H), 1.58 (s, 6H). LCMS: R_(T)=3.76 min, M+H⁺=464

Example 1576,6-dimethyl-4-morpholino-2-(4-(piperidin-1-yl)phenyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine157

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (1)(11 mg, 16 umol) were reacted to give 157. 1H NMR (400 MHz, DMSO) δ 8.21(d, J=8.9 Hz, 2H), 6.96 (d, J=9.0 Hz, 2H), 4.23 (s, 4H), 4.13 (t, J=4.8Hz, 4H), 3.81-3.69 (m, 4H), 3.27-3.21 (m, 4H), 1.61 (m, 6H), 1.58 (s,6H). LCMS: R_(T)=4.13 min, M+H⁺=449

Example 158N-(5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)romoace-2-yl)acetamide 158

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol), N-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)romoace-2-yl)acetamide (0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 158. 1H NMR (400 MHz, DMSO) δ10.64 (s, 1H), 9.23 (d, J=2.2 Hz, 1H), 8.63 (dd, J=8.7, 2.3 Hz, 1H),8.17 (d, J=8.8 Hz, 1H), 4.27 (s, 4H), 4.15 (dd, J=15.2, 5.1 Hz, 4H),3.81-3.71 (m, 4H), 2.12 (s, 3H), 1.59 (s, 6H). LCMS: R_(T)=3.95 min,M+H⁺=424

Example 1595-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)picolinamide159

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinamide (0.31mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (I)(11 mg, 16 umol) were reacted to give 159. 1H NMR (400 MHz, DMSO) δ 9.52(d, J=1.8 Hz, 1H), 8.81 (dd, J=8.2, 2.1 Hz, 1H), 8.14 (d, J=8.2 Hz, 1H),8.12 (s, 1H), 7.69 (s, 1H), 4.29 (s, 4H), 4.18 (m, 4H), 3.83-3.71 (m,4H), 1.60 (s, 6H). LCMS: R_(T)=4.39 min, M+H⁺=410

Example 1606-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)romoace-3-ol 160

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol), 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)romoace-3-ol (0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 160. 1H NMR (400 MHz, DMSO) δ 8.24(m, 2H), 7.24 (dd, J=8.6, 2.9 Hz, 1H), 6.62 (s, 1H), 4.25 (s, 4H), 4.14(d, J=2.5 Hz, 4H), 3.81-3.70 (m, 4H), 1.59 (s, 6H). LCMS: R_(T)=3.73min, M+H⁺=383

Example 161(4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)(4-methylpiperazin-1-yl)methanone161

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),(4-methylpiperazin-1-yl)(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 161. 1H NMR (400 MHz, DMSO) δ 8.43(d, J=8.2 Hz, 2H), 7.47 (d, J=8.2 Hz, 2H), 4.28 (s, 4H), 4.21-4.08 (m,4H), 3.82-3.71 (m, 4H), 3.62 (s, 4H), 2.33 (s, 4H), 2.20 (s, 3H), 1.60(s, 6H). LCMS: R_(T)=3.64 min, M+H⁺=492

Example 162N-cyclopropyl-3-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)benzamide162

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),N-cyclopropyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 162. 1H NMR (400 MHz, DMSO) δ 8.77(s, 1H), 8.53 (d, J=4.0 Hz, 1H), 8.50 (d, J=7.8 Hz, 1H), 7.84 (d, J=7.7Hz, 1H), 7.53 (t, J=7.7 Hz, 1H), 4.28 (s, 4H), 4.17 (dt, J=9.5, 4.4 Hz,4H), 3.84-3.70 (m, 4H), 2.88 (tq, J=7.8, 4.0 Hz, 1H), 0.76-0.67 (m, 2H),0.64-0.54 (m, 2H). LCMS: R_(T)=4.68 min, M+H⁺=449

Example 1635-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)-N,N-dimethylpyrazin-2-amine163

Following General Procedure A,2-chloro-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(102 mg, 0.31 mmol),N,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazin-2-amine(0.31 mmole), andbis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II)(11 mg, 16 umol) were reacted to give 163. 1H NMR (400 MHz, DMSO) δ 9.04(s, 1H), 8.21 (s, 1H), 4.24 (s, 4H), 4.14 (t, J=5.3 Hz, 4H), 3.81-3.66(m, 4H), 3.15 (s, 6H), 1.59 (s, 6H). LCMS: R_(T)=3.93 min, M+H⁺=411

Example 1672-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholino-8,9-dihydropyrazino[2,1-e]purin-6(7H)-one167 Step 1:2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purine-8-carboxylicacid methylamide

A solution of2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purine (0.50 g,1.55 mmol) and N,N,N′,N-tetramethylethylenediamine (0.35 mL, 2.33 mmol)in dry THF (14 mL) was cooled to −78° C. Butyllithium (2.5M in hexanes,1.22 mL, 3.05 mmol) was added dropwise and the dark yellow solution wasstirred at −78° C. for 45 min. N-Succinimidyl N-methylcarbamate (0.4 g,2.33 mmol) was added as a suspension in a small volume of THF and themixture was allowed to warm to room temperature while stirring for 18 h.The reaction mixture was diluted with water, neutralized with 1Mhydrochloric acid and extracted three times with ethyl acetate. Thecombined extracts were dried (Na₂SO₄), filtered and concentrated invacuo. The resulting residue was subjected to flash chromatography(SiO₂, gradient 0-100% ethyl acetate in cyclohexane) to give2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purine-8-carboxylicacid methylamide (104 mg, 18%). LCMS R_(T)=3.26, [M+H]⁺=381/383

Step 2: 2-Chloro-6-morpholin-4-yl-9H-purine-8-carboxylic acidmethylamide

2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purine-8-carboxylicacid methylamide (104 mg, 0.27 mmol) was suspended in methanol (6 mL)and p-toluenesulfonic acid monohydrate (10 mg, 0.05 mmol) was added. Themixture was stirred at room temperature for 68 h, then diluted withwater and neutralized with aqueous sodium bicarbonate. The solidprecipitate was filtered off and dried at 50° C. under vacuum to give2-chloro-6-morpholin-4-yl-9H-purine-8-carboxylic acid methylamide (56mg, 70%). LCMS R_(T)=2.43, [M+H]⁺=297/299

Step 3:3-Chloro-7-methyl-1-morpholin-4-yl-6,7-dihydro-5H-2,4,4b,7,9-pentaaza-fluoren-8-one

A mixture of 2-chloro-6-morpholin-4-yl-9H-purine-8-carboxylic acidmethylamide (56 mg, 0.19 mmol) 1,2-dibromoethane (0.058 mL, 0.68 mmol)and cesium carbonate (0.25 g, 0.76 mmol) in DMF (2 mL) was heated at100° C. for 2 h, then cooled, diluted with water and extracted fivetimes with ethyl acetate. The combined extracts were dried (Na₂SO₄) andconcentrated in vacuo. The resulting residue was triturated twice withdiethyl ether and the solid was dried under vacuum to give3-Chloro-7-methyl-1-morpholin-4-yl-6,7-dihydro-5H-2,4,4b,7,9-pentaaza-fluoren-8-one(47 mg, 77%). LCMS R_(T)=2.36, [M+H]⁺=323/325.

Step 4: A mixture of3-chloro-7-methyl-1-morpholin-4-yl-6,7-dihydro-5H-2,4,4b,7,9-pentaaza-fluoren-8-one(47 mg, 0.15 mmol), 2-aminopyrimidine-5-boronic acid pinacol ester (39mg, 0.18 mmol), and cesium carbonate (131 mg, 0.40 mmol) in 1,4-dioxane(1.5 mL) and water (1.5 mL) was purged with argon.Tetrakis(triphenylphosphine)palladium(0) (9 mg, 0.008 mmol) was added,the mixture was purged with argon again and then heated at 100° C.overnight. The mixture was cooled and diluted with water. Theprecipitate was filtered off, washed with water, and then trituratedwith ethanol. The solid was filtered off and dried (vacuum, 50° C.) togive 167 (15 mg, 26%). LCMS R_(T)=2.47, [M+H]⁺=382. ¹H NMR (DMSO-d,+d1-TFA, 400 MHz): δ 9.38 (2H, s), 4.79-4.06 (4H, v. broad), 4.44 (2H,t, J=6.0 Hz), 3.90 (2H, t, J=6.0 Hz), 3.80 (4H, t, J=4.7 Hz), 3.11 (3H,s).

Example 1685-(8,8-Dimethyl-1-morpholin-4-yl-5,8-dihydro-6H-7-oxa-9-thia-2,4-diaza-fluoren-3-yl)-pyrimidin-2-ylamine168 Step 1: 7-bromothieno[3,2-d]pyrimidine-2,4(1H,3H)-dione

Thieno[3,2-d]pyrimidine-2,4(1H,3H)-dione (10.38 g, 61.72 mmol) wasdissolved in acetic acid (230 mL) and Bromine (11.13 mL, 216 mmol) wasadded. The reaction was heated at 80° C. for 3.5 h. Complete reactionwas confirmed by LCMS. The reaction mixture was poured onto ice waterslowly and filtered off the precipitate which was dried overnight undervacuum to give 7-bromothieno[3,2-d]pyrimidine-2,4(1H,3H)-dione (9.1 g,60% yield)

Step 2: 7-bromo-2,4-dichlorothieno[3,2-d]pyrimidine

7-Bromothieno[3,2-d]pyrimidine-2,4(1H,3H)-dione (9.1 g, 37 mmol) wasdissolved in POCl₃ (140 mL, 1500 mmol) and heated at 110° C. with avigreux condensation column attached for 20 hrs. Complete reactionconfirmed by LCMS. Poured onto ice water slowly and filtered off theprecipitate. The product was purified by silica gel chromatography (0 to100% ethyl acetate/heptanes) on the CombiFlash® (Teledyne Isco Co.) Rfsystem and concentrated in vacuo to give7-bromo-2,4-dichlorothieno[3,2-d]pyrimidine (8.4 g, 80% yield)

Step 3: 4-(7-bromo-2-chlorothieno[3,2-d]pyrimidin-4-yl)morpholine

7-Bromo-2,4-dichlorothieno[3,2-d]pyrimidine (2.9 g, 10.0 mmol) wasdissolved in methanol (100 mL, 2000 mmol) and added morpholine (2 mL, 22mmol) and let the reaction mixture stir for 1.5 h. Complete reactionconfirmed by LCMS. Concentrated in vacuo and diluted with water.Extracted with DCM and concentrated in vacuo again. The product waspurified by silica gel chromatography (0 to 100% ethyl acetate/heptanes)on the CombiFlash® Rf system and concentrated in vacuo to give4-(7-bromo-2-chlorothieno[3,2-d]pyrimidin-4-yl)morpholine (1.2 g, 35%yield). ¹H NMR (400 MHz, CDCl₃) δ 7.78 (s, 1H), 4.01 (m, 4H), 3.85 (m,4H)

Step 4: 4-(2-chloro-7-vinylthieno[3,2-d]pyrimidin-4-yl)morpholine

4-(7-bromo-2-chlorothieno[3,2-d]pyrimidin-4-yl)morpholine (3.55 g, 10.6mmol), (2-Ethenyl)tri-n-butyltin (3.41 mL, 11.7 mmol), Pd(PPh₁)₄(613 mg,0.53 mmol), and 1,4-Dioxane (30 mL, 400 mmol) were combined in a scaledtube and heated at 100° C. 19.5 h. Complete reaction was confirmed byLCMS. Concentrated in vacuo and purified by silica gel chromatography (0to 50% ethyl acetate/heptanes) on the CombiFlash® Rf system andconcentrated in vacuo to give4-(2-chloro-7-vinylthieno[3,2-d]pyrimidin-4-yl)morpholine (1.18 g, 39.5%yield)

Step 5: 2-(2-chloro-4-morpholinothieno[3,2-d]pyrimidin-7-yl)ethanol

4-(2-chloro-7-vinylthieno[3,2-d]pyrimidin-4-yl)morpholine (170 mg, 0.6mmol) was dissolved in tetrahydrofuran (10 mL, 100 mmol) and cooled to0° C. under an atmosphere of nitrogen. 0.5 M 9-BBN in hexanes (3.4 mL, 2mmol) was added and the reaction was allowed to warm up to roomtemperature and stir overnight. LCMS indicated mostly starting materialso the reaction was cooled to 0° C. again and added 0.5 M 9-BBN inhexanes (8.0 mL, 4 mmol) and allowed to warm up to room temperature andstir overnight again. Added 20 M hydrogen peroxide (1.4 mL, 20 mmol)followed by 5 M sodium hydroxide in water (2.4 mL, 10 mmol). Thereaction was diluted with water and extracted with ethyl acetate, driedover magnesium sulfate and concentrated in vacuo and was purified bysilica gel chromatography (0 to 50% ethyl acetate/heptanes) on theCombiFlash®) Rf system and concentrated in vacuo to give2-(2-chloro-4-morpholinothieno[3,2-d]pyrimidin-7-yl)ethanol (110 mg, 61%yield)

Step 6:2-(2-chloro-7-(2-hydroxyethyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)propan-2-ol

2-(2-chloro-4-morpholinothieno[3,2-d]pyrimidin-7-yl)ethanol (70 mg, 0.2mmol) was dissolved in tetrahydrofuran (5 mL, 60 mmol) and cooled to−40° C. under an atmosphere of nitrogen. Added 2.5 M n-BuLi in hexanes(370 mL, 0.93 mmol) and allowed to stir for 1 h. Added acetone (86 uL,1.2 mmol) and again stirred at −40° C. for 5 h. Reaction never went tocompletion and was quenched with saturated ammonium chloride andextracted with ethyl acetate, dried over magnesium sulfate andconcentrated in vacuo to give an unpurified to mixture of startingmaterial and2-(2-chloro-7-(2-hydroxyethyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)propan-2-ol(30 mg)

Step 7:3-Chloro-8,8-dimethyl-1-morpholin-4-yl-5,8-dihydro-6H-7-oxa-9-thia-2,4-diaza-fluorene

Impure2-(2-chloro-7-(2-hydroxyethyl)-4-morpholinothieno[3,2-d]pyrimidin-6-yl)propan-2-ol(30 mg) was dissolved in toluene (5 mL, 50 mmol). Trifluoroacetic acid(0.5 mL, 6 mmol) was added and the reaction mixture was heated at 120°C. for two h. Complete reaction was confirmed by LCMS. Diluted withwater and extracted with ethyl acetate, dried over magnesium sulfate,concentrated in vacuo and was purified by silica gel chromatography (0to 100% ethyl acetate/heptanes) on the CombiFlash) Rf system using anamine column and concentrated in vacuo to give3-Chloro-8,8-dimethyl-1-morpholin-4-yl-5,8-dihydro-6H-7-oxa-9-thia-2,4-diaza-fluorene(10 mg, 10% yield)

Step 8:3-Chloro-8,8-dimethyl-1-morpholin-4-yl-5,8-dihydro-6H-7-oxa-9-thia-2,4-diaza-fluorene(10 mg, 0.03 mmol) was dissolved in acetonitrile (2 mL, 40 mmol) andadded 1 M sodium carbonate in water (2 mL, 2 mmol),5-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)pyrimidin-2-amine (9.0 mg,0.041 mmol), andBis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II)(1.1 mg, 0.001 6 mmol). The reaction was placed on the Biotage microwaveat 120° C. for 15 minutes. The aqueous layer was pipetted off and theorganic layer was concentrated in vacuo and was purified by silica gelchromatography (0 to 100% ethyl acetate/heptanes) on the CombiFlash® Rfsystem using a basic alumina column and concentrated in vacuo to give(85% pure) 168 (2.7 mg, 20% yield). M+1: 399.3. ¹H NMR (400 MHz, CDCl₃)δ 9.30 (s, 2H), 5.34 (brs, 2H), 4.08 (t, 2H), 4.01 (m, 4H), 3.87 (m,4H), 2.95 (t, 2H), 1.62 (s, 6H)

Example 1692-(1H-indazol-4-yl)-4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine169 Step 1:4-morpholino-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine

2-Chloro-4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine,from Examples 139 and 140 (90 mg, 0.0002 mol) and1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole(160 mg, 0.0005 mol) were reacted with Pd(dppf)Cl₂([1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complexwith dichloromethane) and cesium carbonate under microwave Suzukipalladium conditions to give4-morpholino-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(90 mg, 90% yield). LC/MS (ESI+): m/z 530 (M+H)

Step 2:4-Morpholino-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine(100 mg, 0.0002 mol) in MeOH (1 mL) was treated with a catalytic amountof p-toluenesulfonic acid (3 mg, 0.02 mmol). The reaction mixture washeated to 50° C. overnight and was then concentrated under reducepressure. The residue was partitioned between water and EtOAc. Theorganic extracts were washed with water, brine, dried over MgSO₄ andconcentrated to dryness to give 169 (13 g, 16% yield). LC/MS (ESI+): m/z446 (M+H). ¹H NMR (400 MHz, DMSO) δ 13.16 (s, 1H), 8.92 (s, 1H), 8.22(d, J=7.2 Hz, 1H), 7.66 (d, J=8.2 Hz, 1H), 7.47 (t, J=7.8 Hz, 1H), 5.93(q, J=6.8 Hz, 1H), 4.55-4.17 (m, 8H), 3.80 (t, J=4.6 Hz, 4H)

Example 1703-(4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenol170

2-Chloro-4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine,from Examples 139 and 140 (50 mg, 0.00015 mol) and3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (76 mg, 0.00034mol) were reacted with Pd(dppf)Cl₂ and cesium carbonate under microwaveSuzuki palladium conditions to give 170 (10 mg, 15% yield). LC/MS(ESI+): m/z 422 (M+H). ¹H NMR (400 MHz, DMSO) δ 7.96-7.74 (m, 2H), 7.26(t, J=8.1 Hz, 1H), 6.83 (dt, J=23.4, 11.6 Hz, 1H), 5.89 (q, I=6.9 Hz,1H), 4.51-4.08 (m, 8H), 3.77 (t, J=4.6 Hz, 4H), 1.23-0.98 (m, 1H)

Example 1715-(4-((2S,6R)-2,6-dimethylmorpholino)-6,6-dimethyl-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine171 Step 1: 2,6-Dichloro-9-(tetrahydro-pyran-2-yl)-9H-purine

A mixture of 2,6-dichloro-9H-purine (10.0 g, 53 mmol),3,4-dihydro-2H-pyran (9.5 mL, 93 mmol) and p-toluenesulfonic acidmonohydrate (1.0 g, 5.0 mmol) in THF (100 mL) was heated at 100° C. for18 h, then cooled to RT and concentrated in vacuo. The resulting residuewas purified by column chromatography (SiO₂, 0 to 10% ethyl acetate incyclohexane) affording 2,6-Dichloro-9-(tetrahydro-pyran-2-yl)-9H-purineas a cream solid (10.9 g, 75%). ¹H NMR (400 MHz, CDCl₃): δ 8.33 (1H, s),5.77 (1H, dd, J=10.4, 2.4 Hz), 4.19 (1H, m), 3.78 (1H, dt, J=11.6, 2.9Hz), 2.17 (1H, m), 2.09 (1H, m), 1.98 (1H, m), 1.87-1.69 (3H, m).

Step 2:2-[2,6-Dichloro-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-propan-2-ol

BuLi (20 mL, 40.0 mmol, 2 M in pentane) was added dropwise to a solutionof 2,6-dichloro-9-(tetrahydro-pyran-2-yl)-9H-purine (8.0 g, 29.3 mmol)and TMEDA (6.4 mL, 42.4 mmol) in anhydrous THF (100 mL) at −78° C. Theresulting dark solution was stirred at −78° C. for 45 min, then acetone(4 mL, 54.5 mmol) was added and the reaction mixture was stirred at −78°C. for 30 min, then at RT for 30 min. The reaction mixture was quenchedwith water and extracted with ethyl acetate. The combined organicextracts were dried (MgSO₄) and concentrated in vacuo. The resultingresidue was purified by column chromatography (SiO₂, 0 to 20% ethylacetate in cyclohexane) affording2-[2,6-Dichloro-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-propan-2-ol asa dark solid (6.0 g, 62%). ¹H NMR (400 MHz, CDCl₃): δ 6.19 (1H, dd,J=11.2, 2.8 Hz), 4.26 (1H, m), 3.77 (1H, m), 2.87 (1H, m), 2.09 (1H, m),1.90-1.71 (1H, m).

Step 3: 2-(2,6-Dichloro-9H-purin-8-yl)-propan-2-ol

HCl (5 mL, 5 mmol, 1 M aqueous solution) was added to a solution of2-[2,6-dichloro-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-propan-2-ol(6.0 g, 20.66 mmol) in a mixture of DCM (15 mL) and methanol (15 mL) andthe resulting solution stirred at RT for 1 h, then concentrated invacuo. The resulting residue was purified by column chromatography(SiO₂, gradient 0 to 50% methanol in DCM) affording2-(2,6-Dichloro-9H-purin-8-yl)-propan-2-ol as a dark solid (3.38 g,66%). LCMS (method A): R_(T)=2.12 min, [M−H]245/247. ¹H NMR (400 MHz,CDCl₃): δ 1.50 (6H, s)

Step 4:1,3-Dichloro-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene

Cesium carbonate (9.3 g, 28.5 mmol) and 1,2-dibromoethane (4.1 mL, 47.6mmol) were added to a solution of2-(2,6-dichloro-9H-purin-8-yl)-propan-2-ol (3.3 g, 13.36 mmol) in DMF(100 mL) and the reaction mixture was heated at 100° C. for 2 h, thenpartitioned between water and ethyl acetate. The organic extract wasseparated and washed with brine, then dried (MgSO₄) and concentrated invacuo. The resulting residue was purified by column chromatography(SiO₂, 0 to 10 to 20% ethyl acetate in cyclohexane) affording1,3-Dichloro-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluoreneas a yellow solid (1.0 g, 27%). ¹H NMR (400 MHz, CDCl₃): δ 4.26 (2H, dd,J=6, 4 Hz), 4.19 (2H, dd, J=6, 4 Hz).

Step 5:3-Chloro-1-((2R,6S)-2,6-dimethyl-morpholin-4-yl)-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene

A mixture of1,3-dichloro-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene(100 mg, 0.366 mmol), (2R,6S)-2,6-dimethyl-morpholine (84 mg, 0.732mmol) and triethylamine (77 μL, 0.55 mmol) in IMS (2 mL) was heated at140° C. for 20 mins in a microwave reactor, then concentrated in vacuo.The resulting residue was purified by column chromatography (SiO₂, 10%ethyl acetate in cyclohexane) affording3-Chloro-1-((2R,6S)-2,6-dimethyl-morpholin-4-yl)-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluoreneas a white solid (128 mg, 99%). LCMS (method A): R_(T)=3.62 min,[M+H]⁺=352.

Step 6: A mixture of3-chloro-1-((2R,6S)-2,6-dimethyl-morpholin-4-yl)-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene(128 mg, 0.36 mmol),5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrimidin-2-ylamine(121 mg, 0.55 mmol), PdCl₂(PPh₃)₂ (26 mg, 0.036 mmol) and sodiumcarbonate (1 mL, 1.0 mmol, 1M aqueous solution) in acetonitrile (4 mL)was degassed and heated at 120° C. for 30 mins in a microwave reactor,then heated thermally at 100° C. for 18 hours. The reaction mixture wasloaded onto an Isolute® SCX-2 cartridge which was washed with methanoland the product eluted with 2M ammonia in methanol. The basic fractionswere combined and concentrated in vacuo. The resulting residue waspurified by reverse phase HPLC (Phenomenex Gemini 5 μm C18, 0.1% HCO₂Hin water gradient 5-98% acetonitrile) affording 171 as a white solid (10mg, 7%). LCMS (method B): R_(T)=4.14 min, [M+H]⁺=411. ¹H NMR (400 MHz,CDCl₃): δ 9.26 (2H, s), 5.49-5.30 (2H, v. broad s), 5.20 (2H, broad s),4.21 (2H, m), 4.15 (2H, m), 3.75 (2H, m), 2.80 (2H, m), 1.67 (6H, s),1.30 (6H, d, J=6.8 Hz)

Example 1725-(4-(2,2-dimethylmorpholino)-6,6-dimethyl-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine172 Step 1:3-Chloro-1-(2,2-dimethyl-morpholin-4-yl)-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene

A mixture of1,3-dichloro-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene(100 mg, 0.366 mmol), 2,2-dimethyl-morpholine (84 mg, 0.732 mmol) andtriethylamine (77 μL, 0.55 mmol) in IMS (2 mL) was heated at 140° C. for20 mins in a microwave reactor, then concentrated in vacuo. Theresulting residue was purified by column chromatography (SiO₂, 10% ethylacetate in cyclohexane) affording3-Chloro-1-(2,2-dimethyl-morpholin-4-yl)-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluoreneas a white solid (110 mg, 85%). LCMS (method A): R_(T)=3.51 min,[M+H]⁺=352.

Step 2: A mixture of3-chloro-1-(2,2-dimethyl-morpholin-4-yl)-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene(110 mg, 0.31 mmol),5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrimidin-2-ylamine(104 mg, 0.47 mmol), PdCl₂(PPh₃)₂ (22 mg, 0.031 mmol) and sodiumcarbonate (I mL, 1.0 mmol, 1M aqueous solution) in acetonitrile (4 mL)was degassed and heated at 120° C. for 30 mins in a microwave reactor,then heated at 100° C. for 18 hours. The reaction mixture was loadedonto an Isolute® SCX-2 cartridge which was washed with methanol and theproduct eluted with 2M ammonia in methanol. The basic fractions werecombined and concentrated in vacuo. The resulting residue was purifiedby reverse phase HPLC (Phenomenex Gemini 5 μm C18, 0.1% HCO₂H in watergradient 5-98% acetonitrile) affording 172 as a white solid (11 mg, 9%).LCMS (method B): R_(T)=4.00 min, [M+H]⁺=411. ¹H NMR (400 MHz, CDCl₃): δ9.25 (2H, s), 5.20 (2H, broad s), 4.46-4.13 (8H, m), 3.88 (2H, m), 1.66(6H, s), 1.28 (6H, s).

Example 174 5-(4-((1S,4S)-2-oxa-5-azabicyclo[2.2.1I]heptan-5-yl)-6,6-dimethyl-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine174 Step 11:3-Chloro-8,8-dimethyl-1-(1S,4S)-2-oxa-5-aza-bicyclo[2.2.1]hept-5-yl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene

A mixture of1,3-dichloro-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene(100 mg, 0.366 mmol), (1S,4S)-2-oxa-5-aza-bicyclo[2.2.1]heptane (73 mg,0.732 mmol) and triethylamine (77 μL, 0.55 mmol) in IMS (2 mL) washeated at 140° C. for 20 mins in a microwave reactor, then concentratedin vacuo. The resulting residue was purified by column chromatography(SiO₂, 10% ethyl acetate in pentane) affording3-chloro-8,8-dimethyl-1-(1S,4S)-2-oxa-5-aza-bicyclo[2.2.1]hept-5-yl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluoreneas a yellow solid (120 mg, 98%). LCMS (method A): R_(T)=2.81 min,[M+H]⁺=336.

Step 2: A mixture of3-chloro-8,8-dimethyl-1-(1S,4S)-2-oxa-5-aza-bicyclo[2.2.1]hept-5-yl-5,6-dihydro-8H-7-oxa-2,4,4b,9-tetraaza-fluorene(120 mg, 0.36 mmol),5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrimidin-2-ylamine (87mg, 0.39 mmol), Pd(PPh₃)₄(21 mg, 0.018 mmol) and cesium carbonate (163mg, 0.5 mmol) in a mixture of 1,4-dioxane (1.5 mL) and water (0.5 mL)was degassed and heated at 130° C. for 20 mins in a microwave reactor.The reaction mixture was loaded onto an Isolute® SCX-2 cartridge whichwas washed with methanol and the product eluted with 2M ammonia inmethanol. The basic fractions were combined and concentrated in vacuo.The resulting residue was purified by reverse phase HPLC (PhenomenexGemini 5 μm C18, 0.1% HCO₂H in water gradient 5-98% acetonitrile)affording 174 as a white solid (45 mg, 32%). LCMS (method B): R_(T)=3.31min, [M+H]⁺=395. ¹H NMR (400 MHz, DMSO-d₆, 80° C.): δ 9.08 (2H, s), 6.54(2H, broad s), 5.75 (1H, broad s), 4.71 (1H, s), 4.12 (4H, m), 3.87 (1H,dd, J=7.4, 1.4 Hz), 3.79 (2H, s), 3.75 (1H, d, J=7.4 Hz), 1.95 (2H, s),1.59 (6H, d, J=6.7 Hz)

Example 1752-(2-aminopyrimidin-5-yl)-6-methyl-4-morpholino-6,7-dihydropyrazino[2,1-e]purin-8(9H)-one175 Step 1:1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethanone

A solution of2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purine (0.50 g,1.55 mmol) and N,N,N′,N-tetramethylethylenediamine (0.35 mL, 2.33 mmol)in dry THF (14 mL) was cooled to −78° C. n-BuLi (2.5M in hexanes, 1.22mL, 3.05 mmol) was added dropwise and the mixture was stirred at −78° C.for 40 min. N-methyl-N-methoxyacetamide (0.25 mL, 2.33 mmol) was addeddropwise and the mixture was stirred at −78° C. for 1.5 h, then allowedto warm to −30° C. Water was added followed by 1M aqueous HCl and themixture was extracted seven times with ethyl acetate. The combinedorganic extracts were dried (Na₂ SO₄), filtered and concentrated invacuo. The resulting residue was subjected to flash chromatography(SiO2), gradient 0-50% ethyl acetate in cyclohexane to give1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethanone(0.47 g, 83%). LCMS R_(T)=3.57 min, [M+H]⁺=366/368

Step 2:1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethanol

To a stirred suspension ofI-[2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethanone(0.47 g, 1.29 mmol) in ethanol (8 mL) and THF (8 mL) was added sodiumborohydride (49 mg, 1.30 mmol). The reaction mixture was stirred at roomtemperature for 1.5 h, then concentrated in vacuo. The resulting residuewas dissolved in ethyl acetate and aqueous sodium bicarbonate and thephases were separated. The aqueous phase was extracted twice with ethylacetate. The combined organic fractions were dried (Na₂ SO₄), filteredand concentrated in vacuo to give1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethanol(0.50 g, quantitative). LCMS R_(T)=3.20 min, [M+H]⁺=368/370

Step 3:8-(1-Azido-ethyl)-2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purine

1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethanol(0.37 g, 1.01 mmol) was dissolved in anhydrous toluene (5.6 mL) and DMF(0.9 mL) and the solution was cooled in ice. Diphenylphosphoryl azide(0.56 mL, 2.54 mmol) was added, followed by dropwise addition of1,8-diazabicyclo[5.4.0]undec-7-ene (0.37 mL, 2.54 mmol). The reactionmixture was stirred at room temperature for 16 h, then diluted withethyl acetate followed by water and the phases were separated. Theaqueous phase was extracted twice with ethyl acetate and the combinedorganic fractions were dried (Na₂SO₄), filtered and concentrated invacuo. The resulting residue, together with crude product from areaction performed similarly on a smaller scale (0.10 g, 0.27 mmol of1-[2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethanol),was subjected to flash chromatography (SiO2) gradient 0-50% ethylacetate in cyclohexane to give8-(1-Azido-ethyl)-2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purineas two separated pairs of diastereomers (0.25 g and 0.27 g, total 0.52g, 100%). LCMS R_(T)=4.05 min, [M+H]⁺=393/395. LCMS R_(T)=4.16 min,[M+H]⁺=393/395

Step 4:1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethylamine

To a solution of8-(i-azido-ethyl)-2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purine(0.47 g, 1.20 mmol) in THF (13 mL) and water (4 mL) was addedtriphenylphosphine (0.33 g. 1.28 mmol). The reaction mixture was heatedat 70° C. for 2 h, then cooled to RT. Ethyl acetate was added and thephases were separated. The aqueous phase was extracted three times withethyl acetate. The combined organic fractions were dried (Na₂ SO₄),filtered and concentrated in vacuo. The resulting residue, together withcrude product from a reaction performed similarly on a smaller scale(0.05 g, 0.14 mmol of8-(1-azido-ethyl)-2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purine),was subjected to flash chromatography (SiO2) gradient 0-10% methanol inDCM). Eluted material containing title compound and triphenylphosphineoxide was subjected to flash chromatography (SiO2) gradient 0-20%methanol in TBME), and clean material from the two columns was combinedto give1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethylamine(0.41 g, 84%, mixture of two pairs of diastereomers). LCMS R_(T)=2.15min and 2.19 min, [M+H]⁺=367/369

Step 5:2-Bromo-N-{1-[2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethyl}-acetamide

To a solution of1-[2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethylamine(0.25 g, 0.68 mmol) in anhydrous DCM were added romoacetyl bromide (62ILL, 0.74 mmol) and triethylamine (0.13 mL, 0.93 mmol). The mixture wasstirred at RT. After 2 h, another portion of romoacetyl bromide (12 μL)was added and stirring continued for 1.5 h. Water was added, the phaseswere separated and the aqueous phase was extracted twice with DCM. Thecombined organic fractions were dried (Na₂SO₄), filtered andconcentrated in vacuo to give2-Bromo-N-{1-[2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethyl}-acetamide(385 mg, mixture of two pairs of diastereomers), which was used in thenext step without purification. LCMS R_(T)=3.45 min and 3.52 min,[M+H]⁺=487/489/491

Step 6:2-Bromo-N-[1-(2-chloro-6-morpholin-4-yl-9H-purin-8-yl)-ethyl]-acetamide

2-Bromo-N-{1-[2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethyl}-acetamide(crude from step 5, 385 mg) was dissolved in methanol (15 mL) andp-toluenesulfonic acid monohydrate (35 mg) was added. The mixture wasstirred at RT for 16 h. Water was added, aqueous sodium bicarbonate wasadded to give pH 7 and the precipitate was filtered off, washed withwater and dried (in vacuo, 50° C.) to give2-Bromo-N-[1-(2-chloro-6-morpholin-4-yl-9H-purin-8-yl)-ethyl]-acetamide(207 mg, 76%). The aqueous filtrate was extracted three times with ethylacetate. The organic layer was separated and dried (Na₂SO₄), thenconcentrated in vacuo to afford a further crop of less pure2-Bromo-N-[1-(2-chloro-6-morpholin-4-yl-9H-purin-8-yl)-ethyl]-acetamide(39 mg). LCMS R_(T)=2.57 min, [M+H]⁺=403/405/407

Step 7:3-Chloro-8-methyl-1-morpholin-4-yl-7,8-dihydro-2,4,4b,7,9-pentaaza-fluoren-6-one

A mixture of2-bromo-N-[1-(2-chloro-6-morpholin-4-yl-9H-purin-8-yl)-ethyl]-acetamide(275 mg, 0.68 mmol) and cesium carbonate (0.48 g, 1.36 mmol) inanhydrous DMF (10 mL) was stirred at RT for 2 h, then diluted with waterand extracted five times with ethyl acetate. The combined organicextracts were dried (Na₂SO₄), filtered and concentrated in vacuo. Theresulting residue was subjected to flash chromatography (SiO2) gradient0-5% methanol in DCM) to give3-Chloro-8-methyl-1-morpholin-4-yl-7,8-dihydro-2,4,4b,7,9-pentaaza-fluoren-6-one(93 mg, 42%). LCMS R_(T)=2.41 min, [M+H]⁺=323/325

Step 8: A mixture of3-chloro-8-methyl-1-morpholin-4-yl-7,8-dihydro-2,4,4b,7,9-pentaaza-fluoren-6-one(46 mg, 0.14 mmol),), 2-aminopyrimidine-5-boronic acid pinacol ester (78mg, 0.36 mmol), potassium fluoride (46 mg, 0.80 mmol) and Pd(PPh₃)₄(18mg, 0.016 mmol) in anhydrous 1,4-dioxane (5 mL) was heated at 100° C.for 16 h. After cooling to RT, the mixture was diluted with water andthe precipitate formed was filtered off and washed with water. The solidwas triturated with methanol, DCM and finally with acetonitrile to give175 (8 mg, 15%). LCMS R_(T)=2.59 min, [M+H]⁺=382. ¹H NMR (DMSO-d6, 400MHz): δ 9.11 (2H, s), 8.68 (1H, s), 7.05 (2H, s), 4.84 (1H, q, J=6.9Hz), 4.76 (1H, d, J=17.2 Hz), 4 69 (1H, d, J=17.2 Hz), 4.25 (4H, broad),3.75 (4H, t, J=4.5 Hz), 1.56 (3H, d, J=6.9 Hz).

Example 1765-(6,7-dimethyl-4-morpholino-6,7,8,9-tetrahydropyrazino[2,1-e]purin-2-yl)pyrimidin-2-amine176 Step 1:{1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethyl}-carbamicacid tert-butyl ester

To solution of1-[2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethylamine(0.41 g, 1.12 mmol) in anhydrous DCM (10 mL) was added triethylamine(0.17 mL, 1.23 mmol) and di-tert-butyl dicarbamate (0.268 g, 1.23 mmol).The mixture was stirred at RT for 2 h, then washed with 10% aqueouscitric acid. The aqueous phase was extracted three times with DCM, andthe combined organic fractions were dried (Na₂SO₄), filtered andconcentrated in vacuo. The resultant residue was triturated with diethylether, collected by filtration and dried (in vacuo, 50° C.) to give{1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethyl}-carbamicacid tert-butyl ester (0.425 g, 81%, mixture of two pairs ofdiastereomers). LCMS R_(T)=4.06 and 4.13 min, [M+H]⁺=467/469

Step 2:{1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethyl}-methyl-carbamicacid tert-butyl ester

A solution of{1-[2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethyl}-carbamicacid tert-butyl ester (218 mg, 0.47 mmol) in anhydrous THF (15 mL) wascooled to 0° C. Sodium hydride (60% suspension in oil, 22 mg, 0.56 mmol)was added and the mixture was stirred at 0° C. for 30 min. Iodomethane(10 vol. % solution in THF, 0.35 mL, 0.56 mmol) was added and themixture was stirred for 16 h at RT. The reaction mixture was combinedwith another mixture, prepared in a similar manner from 210 mg of thecarbamate starting-material, and diluted with water. After adjustment topH7 by addition of 1 M aqueous HCl and aqueous sodium bicarbonate, themixture was extracted three times with ethyl acetate. The combinedextracts were dried (Na₂SO₄), filtered and concentrated in vacuo, togive a 2:1 mixture of the title compound and carbamate starting-material(0.46 g). This mixture was dissolved in anhydrous THF (20 mL) andtreated with sodium hydride (19 mg) and iodomethane (10 vol. % solutionin THF, 0.29 mL) as before. After addition of more iodomethane (neat,0.050 mL) and stirring for another 8 h, the reaction mixture was dilutedwith water, neutralized and extracted as before. The organic extractswere dried (Na₂SO₄), filtered and concentrated in vacuo. The resultingresidue was subjected to flash chromatography (SiO2) gradient 0-20%ethyl acetate in cyclohexane) to give{1-[2-Chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethyl}-methyl-carbamicacid tert-butyl ester (316 mg, 72%). LCMS R_(T)=4.48 min, [M+H]⁺=481/483

Step 3: [1-(2-Chloro-6-morpholin-4-yl-9H-purin-8-yl)-ethyl]-methyl-amine

A mixture of{1-[2-chloro-6-morpholin-4-yl-9-(tetrahydro-pyran-2-yl)-9H-purin-8-yl]-ethyl}-methyl-carbamicacid tert-butyl ester (316 mg, 0.66 mmol) and p-toluenesulfonic acidmonohydrate (30 mg) in methanol was stirred at room temperature for 3 h,then allowed to stand for 56 h. The reaction mixture was concentrated invacuo to a small volume, then DCM (3 mL) and trifluoroacetic acid (3 mL)were added and the reaction mixture was stirred for 4.75 h at RT.Another portion of trifluoroacetic acid (3 mL) was added and stirringwas continued for 2 h. The mixture was concentrated in vacuo and theresulting residue was triturated three times with diethyl ether. Theresulting solid was partitioned between a 10% solution of methanol inDCM and aqueous sodium bicarbonate. The phases were separated, and theaqueous phase was extracted four times with a 10% solution of methanolin DCM. The combined organic fractions were dried (Na₂SO₄), filtered andconcentrated in vacuo to give[1-(2-Chloro-6-morpholin-4-yl-9H-purin-8-yl)-ethyl]-methyl-amine (0.19g, 97%). LCMS R_(T)=1.68 min, [M+H]⁺=297

Step 4:3-Chloro-7,8-dimethyl-1-morpholin-4-yl-5,6,7,8-tetrahydro-2,4,4b,7,9-pentaaza-fluorene

A mixture of:[1-(2-chloro-6-morpholin-4-yl-9H-purin-8-yl)-ethyl]-methyl-amine (95 mg,0.32 mmol), cesium carbonate (652 mg, 2 mmol) and 1,2-dibromoethane(0.030 mL, 0.35 mmol) in DMF (5 mL) was stirred at RT for 4 h. Anotherportion of 1,2-dibromoethane (0.030 mL, 0.35 mmol) was added andstirring was continued for 20 h. The reaction mixture was diluted withwater and extracted seven times with ether. The combined organicextracts were dried (Na₂SO₄), filtered and concentrated in vacuo. Theresultant residue was subjected to flash chromatography (SiO2) gradient0-4% methanol in DCM) to give3-Chloro-7,8-dimethyl-1-morpholin-4-yl-5,6,7,8-tetrahydro-2,4,4b,7,9-pentaaza-fluorene(76 mg, 74%). LCMS R_(T)=1.82 min, [M+H]⁺=323/325

Step 5: A mixture of3-chloro-7,8-dimethyl-1-morpholin-4-yl-5,6,7,8-tetrahydro-2,4,4b,7,9-pentaaza-fluorene(70 mg, 0.22 mmol), 2-aminopyrimidine-5-boronic acid pinacol ester (57mg, 0.26 mmol) and cesium carbonate (216 mg, 0.66 mmol) in 1,4-dioxane(2.5 mL) and water (2.5 mL) was purged with argon. Pd(PPh₃)₄ (12 mg,0.011 mmol) was added, the mixture was purged with argon again and thenheated at 100° C. for 16 h. Further portions of the boronate ester (29mg) and Pd(PPh₃)₄(6 mg) were added and heating was continued for 5.5 h.The mixture was diluted with water and extracted five times with ethylacetate. The combined organic extracts were dried (Na₂SO₄), filtered andconcentrated in vacuo. The resultant residue was subjected to flashchromatography (SiO2) gradient 0-10% methanol in DCM) to give 176 (58mg, 69%). LCMS R_(T)=2.11 min, [M+H]⁺=382. ¹H NMR (DMSO-d₆, 400 MHz: 69.08 (2H, s), 7.01 (2H, s), 4.24 (4H, broad), 4.20 (1H, m), 4.00 (1H,m), 3.75 (4H, t, J=4.8 Hz), 3.55 (1H, q, J=6.6 Hz), 3.21 (1H, m), 2.78(1H, m), 2.43 (3H, s), 1.50 (3H, d, J=6.6 Hz).

Example 1775-(8,8-Dimethyl-1-morpholin-4-yl-5,6-dihydro-8H-7-oxa-2,4,4b-triaza-fluoren-3-yl)-pyrimidin-2-ylamine177 Step 1: 7-Benzenesulfonyl-2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine

A solution of 2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine (1 g, 5.3 mmol)in anhydrous THF (5 mL) was added to a suspension of sodium hydride (234mg, 5.83 mmol, 60% dispersion in mineral oil) in anhydrous THF (15 mL)at 0° C. The resulting mixture was stirred at 0° C. for 45 ruin, andbenzenesulfonyl chloride (1.12 g, 6.36 mmol) was added dropwise. Thereaction mixture was allowed to warm to ambient temperature and stirredat RT for 1 h. The reaction mixture was quenched with saturated aqueousammonium chloride and extracted with ethyl acetate. The combined organicextracts were dried (Na₂SO₄) and concentrated in vacuo. The resultingresidue was triturated with cyclohexane affording7-Benzenesulfonyl-2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine as a yellowsolid (1.52 g, 87%). ¹H NMR (400 MHz, CDCl₃): δ 8.24 (2H, m), 7.76 (1H,d, J=4.0 Hz), 7.69 (1H, m), 7.58 (2H, m), 6.69 (1H, d, J=4.0 Hz).

Step 2: 2-(2,4-Dichloro-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-propan-2-ol

Lithium diisopropylamide (2 mL, 4.0 mmol, 2 M in THF) was added dropwiseto a solution of7-benzenesulfonyl-2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine (656 mg, 2.0mmol) in anhydrous THF (15 mL) at −78° C. The resulting solution wasstirred at −78° C. for 90 min, then acetone (0.4 mL, 5.5 mmol) was addedand the reaction mixture was stirred at −78° C. for 30 min. The reactionmixture was quenched with saturated aqueous ammonium chloride andextracted with ethyl acetate. The combined organic extracts were dried(MgSO₄) and concentrated in vacuo affording2-(7-benzenesulfonyl-2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-propan-2-ol.To a solution of2-(7-benzenesulfonyl-2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-propan-2-ol(2 mmol) in a mixture of isopropyl alcohol (11 mL) and water (3 mL) wasadded sodium hydroxide (6 mL, 36 mmol, 6M aqueous solution). Theresulting mixture was stirred at RT for 2 h, then concentrated in vacuo.The resulting residue was purified by column chromatography (SiO₂,gradient 0 to 40% ethyl acetate in cyclohexane) affording2-(2,4-Dichloro-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-propan-2-ol (304 mg,64%). LCMS (method A): R_(T)=2.70 min, [M]⁻=244/246.

Step 3:3-Chloro-8,8-dimethyl-1-morpholin-4-yl-5,6-dihydro-8H-7-oxa-2,4,4b-triaza-fluorene

Cesium carbonate (1.2 g, 3.7 mmol) and 1,2-dibromoethane (316 μL, 3.7mmol) were added to a solution of2-(2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-propan-2-ol (304 mg,1.23 mmol) in DMF (4 mL) and the reaction mixture was heated at 100° C.for 45 min, then partitioned between water and ethyl acetate. Theorganic extract was separated and washed with brine, then dried (Na₂SO₄)and concentrated in vacuo affording1,3-dichloro-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b-triaza-fluorene. Amixture of1,3-dichloro-8,8-dimethyl-5,6-dihydro-8H-7-oxa-2,4,4b-triaza-fluorene(1.23 mmol), morpholine (236 μL, 2.69 mmol) and triethylamine (342 μL,2.46 mmol) in IMS (3 mL) was heated at reflux for 3 h, then concentratedin vacuo. The resulting residue was purified by column chromatography(SiO₂, gradient 0 to 40% ethyl acetate in cyclohexane) affording3-Chloro-8,8-dimethyl-1-morpholin-4-yl-5,6-dihydro-8H-7-oxa-2,4,4b-triaza-fluorene(159 mg, 40%). LCMS (method A): R_(T)=3.13 min, [M+H]⁺=323.

Step 4: A mixture of3-Chloro-8,8-dimethyl-1-morpholin-4-yl-5,6-dihydro-8H-7-oxa-2,4,4b-triaza-fluorene(75 mg, 0.23 mmol),5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyrimidin-2-ylamine(115 mg, 0.52 mmol),bis(di-tert-butyl(4-dimethylaminophenyl)-phosphine)dichloropalladium(II)(25 mg, 0.035 mmol) and sodium carbonate (1 mL, 1.0 mmol, 1M aqueoussolution) in acetonitrile (3 mL) was degassed and heated at 150° C. for30 mins in a microwave reactor, then concentrated in vacuo. Theresulting residue was purified by column chromatography (SiO₂, gradient0 to 75% ethyl acetate in cyclohexane) followed by reverse phase HPLC(Phenomenex Gemini 5 μm C18, 0.1% HCO₂H in water on a gradientacetonitrile 5-98%) affording 177 as a off-white solid (13 mg, 15%).LCMS (method B): R_(T)=3.50 min, [M+H]⁺=382. ¹H NMR (400 MHz, CDCl₃): δ9.27 (2H, s), 6.12 (1H, s), 5.27 (2H, broad s), 4.21 (2H, m), 4.14 (2H,m), 3.97 (4H, m), 3.88 (4H, m), 1.62 (6H, s)

Example 901 p110α (Alpha) PI3K Binding Assay

Binding Assays: Initial polarization experiments were performed on anAnalyst HT® 96-384 (Molecular Devices Corp, Sunnyvale, Calif.). Samplesfor fluorescence polarization affinity measurements were prepared byaddition of 1:3 serial dilutions of p110 alpha PI3K (Upstate CellSignaling Solutions, Charlottesville, Va.) starting at a finalconcentration of 20 ug/mL in polarization buffer (10 mM tris pH 7.5, 50mM NaCl, 4 mM MgCl₂, 0.05% Chaps, and 1 mM DTT) to 10 mM PIP₂(Echelon-Inc., Salt Lake City, Utah) final concentration. After anincubation time of 30 minutes at room temperature, the reactions werestopped by the addition of GRP-1 and PIP3-TAMRA probe (Echelon-Inc.,Salt Lake City, Utah) 100 nM and 5 nM final concentrations respectively.Read with standard cut-off filters for the rhodamine fluorophore(λex=530 nm; λem=590 nm) in 384-well black low volume Proxiplates®(PerkinElmer, Wellesley, Mass.) Fluorescence polarization values wereplotted as a function of the protein concentration, and the EC₅₀ valueswere obtained by fitting the data to a 4-parameter equation usingKalcidaGraph® software (Synergy software, Reading, Pa.). This experimentalso establishes the appropriate protein concentration to use insubsequent competition experiments with inhibitors.

Inhibitor IC₅₀ values were determined by addition of the 0.04 mg/mL p110alpha PI3K (final concentration) combined with PIP₂ (10 mM finalconcentration) to wells containing 1:3 serial dilutions of theantagonists in a final concentration of 25 mM ATP (Cell SignalingTechnology, Inc., Danvers, Mass.) in the polarization buffer. After anincubation time of 30 minutes at room temperature, the reactions werestopped by the addition of GRP-1 and PIP3-TAMRA probe (Echelon-Inc.,Salt Lake City, Utah) 100 nM and 5 nM final concentrations respectively.Read with standard cut-off filters for the rhodamine fluorophore(λex=530 nm; λem=590 nm) in 384-well black low volume PROXIPLATES®(PerkinElmer, Wellesley, Mass.) Fluorescence polarization values wereplotted as a function of the antagonist concentration, and the IC₅₀values were obtained by fitting the data to a 4-parameter equation inAssay Explorer® software (MDL, San Ramon, Calif.).

Alternatively, inhibition of PI3K was determined in a radiometric assayusing purified, recombinant enzyme and ATP at a concentration of 1 μM.The Formula I compound was serially diluted in 100% DMSO. The kinasereaction was incubated for 1 hr at room temperature, and the reactionwas terminated by the addition of PBS. IC₅₀ values were subsequentlydetermined using sigmoidal dose-response curve fit (variable slope).

Example 902 In Vitro Cell Proliferation Assay

Efficacy of Formula I compounds was measured by a cell proliferationassay employing the following protocol (Promega Corp. Technical BulletinTB288; Mendoza et al (2002) Cancer Res. 62:5485-5488):

1. An aliquot of 100 μl of cell culture containing about 10⁴ cells (PC3,Detroit562, or MDAMB361.1) in medium was deposited in each well of a384-well, opaque-walled plate.

2. Control wells were prepared containing medium and without cells.

3. The compound was added to the experimental wells and incubated for3-5 days.

4. The plates were equilibrated to room temperature for approximately 30minutes.

5. A volume of CellTiter-Glo® Reagent equal to the volume of cellculture medium present in each well was added.

6. The contents were mixed for 2 minutes on an orbital shaker to inducecell lysis.

7. The plate was incubated at room temperature for 10 minutes tostabilize the luminescence signal.

8. Luminescence was recorded and reported in graphs as RLU=relativeluminescence units.

Alternatively, cells were seeded at optimal density in a 96 well plateand incubated for 4 days in the presence of test compound. Alamar Blue™was subsequently added to the assay medium, and cells were incubated for6 hr before reading at 544 nm excitation, 590 nm emission. EC₅₀ valueswere calculated using a sigmoidal dose response curve fit. The term EC₅₀refers to the half maximal effective concentration and is theconcentration at which a drug induces a response halfway between thebaseline and maximum after some specified exposure time. It is commonlyused as a measure of drug potency.

The anti-proliferative effects of Formula I exemplary compounds weremeasured by the CellTiter-Glo® Assay against various tumor cell lines,including the following:

Cell line: MDA-MB-361.1 Cell line: PC3 Tissue type: breast Tissue type:prostate Mutation Status: PI3K Mutation Status: PTEN Compound No. EC50(μmole) EC50 (μmole) 101  0.315  0.208 102  0.081 2   103  0.269  0.44115  0.958 1.8 121  0.36 — 122 3.2 2   138 10+   10+   139  0.424  0.697148 10+   10+   163 10+   10+   165 1.2 1.3 168  0.237  0.215 169 4.46.9 170  0.92 1.1 171 3.1 3.8 172 1.5 1.4 174 3.4 3.6 176  0.889 1.9 177 0.094  0.185

Example 903 Caco-2 Permeability

Caco-2 cells are seeded onto Millipore Multiscreen®) plates at 1×10⁵cells/cm², and cultured for 20 days. Assessment of compound permeabilityis subsequently conducted. The compounds are applied to the apicalsurface (A) of cell monolayers and compound permeation into thebasolateral (B) compartment is measured. This is performed in thereverse direction (B-A) to investigate active transport. A permeabilitycoefficient value, P_(app), for each compound, a measure of the rate ofpermeation of the compound across the membrane, is calculated. Compoundsare grouped into low (P_(app), </=1.0×10⁶ cm/s) or high(P_(app)>/=1.0×10⁶ cm/s) absorption potential based on comparison withcontrol compounds with established human absorption.

For assessment of a compound's ability to undergo active efflux, theratio of basolateral (B) to apical (A) transport compared with A to Bwas determined. Values of B-A/A-B>/=1.0 indicate the occurrence ofactive cellular efflux.

Example 904 Hepatocyte Clearance

Suspensions of cryopreserved human hepatocytes are used. Incubations areperformed at compound concentration of 1 mM or 3 μM at a cell density of0.5×10⁶ viable cells/mL. The final DMSO concentration in the incubationis about 0.25%. Control incubations are also performed in the absence ofcells to reveal any non-enzymatic degradation. Duplicate samples (50 μL)are removed from the incubation mixture at 0, 5, 10, 20, 40 and 60minutes (control sample at 60 minutes only) and added to methanolcontaining internal standard (100 μL)—to terminate the reaction.Tolbutamide, 7-hydroxycoumarin, and testosterone may be used as controlcompounds. Samples are centrifuged and the supernatants at each timepoint pooled for analysis by LC-MSMS. From a plot of ln peak area ratio(parent compound peak area/internal standard peak area) against time,intrinsic clearance (CL_(int)) is calculated as follows: CL_(int)(μl/min/million cells)=V×k, where k is the elimination rate constant,obtained from the gradient of in concentration plotted against time; Vis a volume term derived from the incubation volume and is expressed asuL 10⁶ cells⁻¹.

Example 905 Cytochrome P450 Inhibition

Formula I compounds may be screened against CYP450 targets (1A2, 2C9,2C19, 2D6, 3A4) at about 10 concentrations in duplicate, with a topconcentration of about 100 uM. Standard inhibitors (furafylline,sulfaphenazole, tranylcypromine, quinidine, ketoconazole) may be used ascontrols. Plates may be read using a BMG LabTechnologies PolarStar™ influorescence mode.

Example 906 Cytochrome P450 Induction

Freshly isolated human hepatocytes from a single donor may be culturedfor about 48 hr prior to addition of Formula I compound at threeconcentrations and incubated for 72 hr. Probe substrates for CYP3A4 andCYP1A2 are added for 30 minutes and 1 hr before the end of theincubation. At 72 hr, cells and media are removed and the extent ofmetabolism of each probe substrate quantified by LC-MS/MS. Theexperiment is controlled by using inducers of the individual P450sincubated at one concentration in triplicate.

Example 907 Plasma Protein Binding

Solutions of Formula I compound (5 um, 0.5% final DMSO concentration)are prepared in buffer and 10% plasma (v/v in buffer). A 96 well HTdialysis plate is assembled so that each well is divided in two by asemi-permeable cellulose membrane. The buffer solution is added to oneside of the membrane and the plasma solution to the other side;incubations are then conducted at 37° C. over 2 hr in triplicate. Thecells are subsequently emptied, and the solutions for each batch ofcompounds are combined into two groups (plasma-free andplasma-containing) then analyzed by LC-MSMS using two sets ofcalibration standards for plasma-free (6 points) and plasma-containingsolutions (7 points). The fraction unbound value for the compound iscalculated.

Example 908 hERG Channel Blockage

Formula I compounds are evaluated for ability to modulate rubidiumefflux from HEK-294 cells stably expressing hERG potassium channelsusing established flux methodology. Cells are prepared in mediumcontaining RbCl, plated into 96-well plates and grown overnight to formmonolayers. The efflux experiment is initiated by aspirating the mediaand washing each well with 3×100 μL of pre-incubation buffer (containinglow [K⁺]) at room temperature. Following the final aspiration, 50 μL ofworking stock (2×) compound is added to each well and incubated at roomtemperature for 10 minutes. Stimulation buffer 50 μL (containing high[K+]) is then added to each well giving the final test compoundconcentrations. Cell plates are then incubated at room temperature for afurther 10 minutes. Supernatant 80 μL from each well is then transferredto equivalent wells of a 96-well plate and analyzed via atomic emissionspectroscopy. The compound is screened as 10 pt duplicate IC₅₀ curves,n=2, from a top concentration of 100 μM.

Example 909 In Vivo Tumor Xenograft

NCR nude mice (Taconic Farms, Ind.) were inoculated subcutaneously inthe right lateral thorax with 5 million U-87 MG Merchant (an in-housevariant derived from U-87 MG cells from ATCC, Manassas, Va.) cells inHBSS/Matrigel (1:1, v/v). Mice bearing tumor xenografts were dosed dailyorally by gavage for <28 days with drug or vehicle after being separatedinto different dose groups of similarly sized tumors. Tumor sizes wererecorded at least twice weekly over the course of the study. Mouse bodyweights were also recorded at least twice weekly, and the mice wereobserved daily. Tumor volume was measured in two perpendiculardimensions (length and width) using Ultra Cal-IV calipers (Model54-10-111; Fred V. Fowler Co., Inc.; Newton, Mass.) and analyzed usingExcel v. 11.2 (Microsoft Corporation; Redmond, Wash.). Tumor inhibitiongraphs were plotted using GraphPad Prism™, Version 5.0c (GraphPadSoftware, Inc.; La Jolla, Calif.). The tumor volume was calculated withformula: Tumor size (mm³)=(longer measurement×shorter measurement)×0.5

Animal body weights were measured using an Adventurer Pro™ AV812 scale(Ohaus Corporation; Pine Brook, N.J.). Graphs were generated usingGraphPad Prism™, Version 5.0c. Percent weight change was calculatedusing formula: individual percent weight change=((new weight/initialweight)−1)×100.

Mice whose tumor volume exceeded 2000 mm³ or whose body weight lossexceeded 20% of their starting weight were euthanized according toregulatory guidance.

The percent tumor growth inhibition (% TGI) at the end of study (EOS)was calculated using formula:

% TGI=(1−[(AUC!Day)_(Treatment)÷(AUC/Day)_(Control)])×100, where AUC/Dayis the area under the fitted tumor growth curve on the natural scaledivided by the number of study days. Log 2(tumor volume) growth traceswere fitted to each dose group with restricted cubic splines for thefixed time and dose effect in each group. Fitting was done via a linearmixed effects model, using the R package ‘nlme’, version 3.1-97 (11) inR version 2.12.0 (R Development Core Team 2008; R Foundation forStatistical Computing; Vienna, Austria).

A partial response (PR) was defined as a >50% reduction in startingtumor volume that never became a complete response (CR) at any day ofthe study. CR was defined as a 100% reduction in starting tumor volumeat any day of the study. Study tumor incidence (STI) reflected thenumber of animals in a group with a measureable tumor for their lasttumor volume measurement.

Linear mixed effect analysis was also employed to model the percentchange in body weight over time and in response to dose.

Example 910 Phospho AKT Induction Assay

In a 6-well tissue culture plate cells were seeded at 5×10⁵ cells perwell overnight. Cells were treated with an EC₀ of the Formula Icompound. Following treatment, cells were washed once with cold PBS andlysed in IX Cell Extraction Buffer from Biosource (Carlsbad, Calif.)supplemented with protcase inhibitors (Roche, Mannheim, Germany), 1 mMPMSF, and Phosphatase Inhibitor Cocktails 1 and 2 from Sigma (St. Louis,Mo.). Determination of protein concentration was performed using thePierce BCA Protein Assay Kit (Rockford, Ill.). Levels of pAkt (Ser⁴⁷³)and total Akt were assessed using bead kits from Biosource (Carlsbad,Calif.) and the Luminex™ Bio-Plex system (Bio-Rad, Hercules, Calif.).

Example 911 Blood-Brain Barrier Activity/Penetrant Assay MDCK1-MDR1 andMDCK11-Bcrp1 Assays

Madin-Darby canine kidney (MDCK) cells heterologously expressing eitherhuman Pgp or mouse Bcrp1 were used to determine whether the compoundswere substrate of these transporters, and thus assess the potential forblood-brain barrier permeation. MDR1-MDCK1 cells were licensed from theNCI (National Cancer Institute, Bethesda, Md.) while Bcrp1-MDCK11 cellswere obtained from the Netherlands Cancer Institutes (Amsterdam, TheNetherlands). Cells were seeded on 24-well Millipore filter plates 4days prior to use (polyester membrane, 1 μM pore size; Millipore;Billerica, Mass.) at a seeding density of 1.3×10⁵ cells/mL. Compoundswere tested at 5 μM in the apical to basolateral (A-B) and basolateralto apical (B-A) directions. The compounds were dissolved in transportbuffer consisting of Hank's balanced salt solution (HBSS) with 10 mMHEPES (Invitrogen Corporation, Grand Island, N.Y.). Lucifer Yellow(Sigma-Aldrich, St. Louis, Mo.) was used as the paracellular marker. Theapparent permeability (P_(app)) in the A-B and B-A directions wascalculated after a 2-hour incubation using the following equation:

P _(app)=(dQ/dt)×1/C ₀×1/A

where dQ/dt is the rate of compound appearance in the receivercompartment, C₀ is the concentration in the donor compartment and A isthe surface area of the insert. The Efflux Ratio, defined asP_(app(B-A))/P_(app A-B)), was used to assess the extent of activeefflux undergone by the compounds with the transporter tested(P-glycoprotein or bcrp1). The compounds were analyzed by LC-MS/MS

Example 912 Determination of Compound Concentration in the Brain

Brains were collected at 1 and 6 hours post-dose from 3 differentanimals at each time point, rinsed with ice-cold saline, weighed andstore at −80° C. until analysis. For compound quantitation, mouse brainswere homogenized in 3 volumes of water. The homogenates were extractedby protein precipitation with acetonitrile containing the internalstandard. LC-MS/MS analysis was conducted. Brain homogenatesconcentrations were converted to brain concentrations for thecalculations of brain-to-plasma ratios.

Example 913 Measurement of the Modulation of the PI3K Pathway in theBrain

For analysis of PI3K pathway modulation, cell extraction buffer(Invitrogen, Camarillo, Calif.) containing 10 mM Tris pH 7.4, 100 mMNaCl, 1 mM EDTA, 1 mM EGTA, 1 mM NaF, 20 mM Na₄P₂O₇, 2 mM Na₃VO₄, 1%Triton X-100, 10% glycerol, 0.1% SDS, and 0.5% deoxycholate wassupplemented with phosphatase, protease inhibitors (Sigma, St. Louis,Mo.) and 1 mM PMSF and added to frozen brain biopsies. Brains collectedat 1 and 6 hrs post-dose were homogenized with a small pestle (KonteGlass Company, Vineland, N.J.), sonicated briefly on ice, andcentrifuged at 20,000 g for 20 minutes at 4° C. Protein concentrationwas determined using BCA protein assay (Pierce, Rockford, Ill.).Proteins were separated by electrophoresis and transferred to NuPagenitrocellulose membranes (Invitrogen, Camarillo, Calif.). Licor Odyssey™Infrared detection system (Licor, Lincoln, Nebr.) was used to assess andquantify protein expression. PI3K pathway markers were evaluated byimmunoblotting using antibodies against pAkt^(scr473) and total Akt(Invitrogen, Camarillo, Calif. and Cell Signaling, Danvers, Mass.).

Example 914 Brain Tumor In Vivo Efficacy Assay

CD-1 Nude mice (Charles River Laboratories, Hollister, Calif.) wereinoculated intracranially under stereotactic surgery with GS-2 (humanglioblastoma muliforme) cells engineered in-house to express luciferasein HBSS. Mice with confirmed brain xenografts by magnetic resonanceimaging (MRI) at four weeks post cell inoculation were dosed once dailyorally by gavage for 28 days with drug or vehicle after being separatedinto groups of similarly sized tumors. MRI (4.7 T, Varian, Inc., PaloAlto, Calif.) was repeated at the end of the 28-day dosing period toassess response to treatment.

Mouse body weights were recorded at least twice weekly, and the micewere observed daily. Animal body weights were measured using anAdventurer Pro™ AV812 scale (Ohaus Corporation; Pine Brook, N.J.).Graphs were generated using GraphPad Prism, Version 5.0c. Percent weightchange was calculated using formula: individual percent weightchange=((new weight/initial weight)−1)×100. Mice whose body weight lossexceeded 20% of their starting weight were euthanized according toregulatory guidance.

The tumor volume change was modeled as linear over the two times atwhich each animal was imaged. A linear mixed effects model was fitted tothese data using the R package ‘nlme’, version 3.1-97 in R version2.12.0 (R Development Core Team 2010; R Foundation for StatisticalComputing; Vienna, Austria). A mixed effect model takes into accountrepeated measures on individual mice over time and handles theintra-mouse correlation appropriately. Linear mixed effect analysis wasalso employed to model the percent change in body weight over time.

Plasma and brain samples were collected at 2 and 8 hours postadministration of the final treatment for pharmacokinetic (PK),pharmacodynamic (PD), and/or immunohistochemical (IHC) analysis.

Example 915 In Vivo Tumor Xenograft PK/PD study

NCR nude mice (Taconic Farms, Ind.) were inoculated subcutaneously inthe right lateral thorax with 5 million U-87 MG Merchant (an in-housevariant derived from U-87 MG cells from ATCC, Manassas, Va.) cells inHBSS/Matrigel™, BD Biosciences (1:1, v/v). Mice bearing tumorxenografts >600 mm³ were dosed once with drug or vehicle after beingseparated into groups of similarly sized tumors. Plasma, subcutaneoustumor xenograft, skeletal muscle, and brain samples were collected at1,4, 12, and 24 hours post treatment administration for PK, PD, and/orIC analysis.

The words “comprise,” “comprising,” “include,” “including,” and“includes” when used in this specification and in the following claimsare intended to specify the presence of stated features, integers,components, or steps, but they do not preclude the presence or additionof one or more other features, integers, components, steps, or groupsthereof.

We claim:
 1. A compound selected from Formula I:

and stereoisomers, geometric isomers, tautomers, and pharmaceutically acceptable salts thereof, wherein: the dashed lines indicate an optional double bond, and at least one dashed line is a double bond; X¹ is S, O, N, NR^(a), CR¹, C(R¹)₂, or —C(R¹)₂O—; X² is C, CR² or N; X³ is C, CR³ or N; A is a 5, 6, or 7-membered carbocyclyl or heterocyclyl ring fused to X² and X³, optionally substituted with one or more R⁵ groups; R^(a) is H, C₁-C₁₂ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-C(═O)—(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-(C₆-C₂₀ aryl), and —(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl), where alkyl, alkenyl, alkynyl, alkylene, carbocyclyl, heterocyclyl, aryl, and heteroaryl are optionally substituted with one or more groups independently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —C(CH₃)₃, —CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH, —CH₂OCH₃, —CN, —CH₂F, —CHF₂, —CF₃, —CO₂H, —COCH₃, —COC(CH₃)₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —C(CH₃)₂CONH₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCOCH₃, —NHS(O)₂CH₃, —N(CH₃)C(CH₃)₂CONH₂, —N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —S(O)₂CH₃, cyclopropyl, cyclobutyl, oxetanyl, morpholino, and 1,1-dioxo-thiopyran-4-yl; R¹, R², and R³ are independently selected from H, F, Cl, Br, I, —CH₃, —CH₂CH₃, —C(CH₃)₃, —CH₂OH, —CH₂CH₂OOH, —C(CH₃)₂OH, —CH₂OCH₃, —CN, —CF₃, —CO₂H, —COCH₃, —COC(CH₃)₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —C(CH₃)₂CONH₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCOCH₃, —NHS(O)₂CH₃, —N(CH₃)C(CH₃)₂CONH₂, —N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —S(O)₂CH₃, cyclopropyl, cyclobutyl, oxetanyl, morpholino, and 1,1-dioxo-thiopyran-4-yl; R⁴ is selected from C₆-C₂₀ aryl, C₂-C₂₀ heterocyclyl and C₁-C₂₀ heteroaryl, each of which are optionally substituted with one or more groups R⁶ groups independently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —CH₂CH(CH₃)₂, —CH₂CH₃, —CH₂CN, —CN, —CF₃, —CH₂OH, —CO₂H, —CONH₂, —CONH(CH₃), —CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —NHCOCH₃, —OH, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —SH, —NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH₃, —NHC(═O)NHCH(CH₃)₂, —NHS(O)₂CH₃, —N(CH₃)C(═O)OC(CH₃)₃, —S(O)₂CH₃, benzyl, benzyloxy, morpholinyl, morpholinomethyl, and 4-methylpiperazin-1-yl; and R⁵ is independently selected from C₁-C₁₂ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₁₂ alkylene)-(C₃-C₁₂ carbocyclyl), —(C₁-C₁₂ alkylene)-(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-C(═O)—(C₂-C₂₀ heterocyclyl), —(C₁-C₁₂ alkylene)-(C₆-C₂₀ aryl), and —(C₁-C₁₂ alkylene)-(C₁-C₂₀ heteroaryl); or two geminal R⁵ groups form a 3, 4, 5, or 6-membered carbocyclyl or heterocyclyl ring, where alkyl, alkenyl, alkynyl, alkylene, carbocyclyl, heterocyclyl, aryl, and heteroaryl are optionally substituted with one or more groups independently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —C(CH₃)₃, —CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH, —CH₂OCH₃, —CN, —CH₂F, —CHF₂, —CF₃, —CO₂H, —COCH₃, —COC(CH₃)₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —C(CH₃)₂CONH₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCOCH₃, —NHS(O)₂CH₃, —N(CH₃)C(CH₃)₂CONH₂, —N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —S(O)₂CH₃, cyclopropyl, cyclobutyl, oxetanyl, morpholino, and 1,1-dioxo-thiopyran-4-yl; mor is selected from:

optionally substituted with one or more R⁷ groups independently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, —CH₂OCH₃, —CHF₂, —CN, —CF₃, —CH₂OH, —CH₂OCH₃, —CH₂CH₂OH, —CH₂C(CH₃)₂OH, —CH(CH₃)OH, —CH(CH₂CH₃)OH, —CH₂CH(OH)CH₃, —C(CH₃)₂OH—, —C(CH₃)₂OCH₃, —CH(CH₃)F, —C(CH₃)F₂, —CH(CH₂CH₃)F, —C(CH₂CH₃)₂F, —CO₂H, —CONH₂, —CON(CH₂CH₃)₂, —COCH₃, —CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCH₂CH₃, —NHCH(CH₃)₂, —NHCH₂CH₂OH, —NHCH₂CH₂OCH₃, —NHCOCH₃, —NHCOCH₂CH₃, —NHCOCH₂OH, —NHS(O)₂CH₃, —N(CH₃)S(O)₂CH₃, ═O, —OH, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —SH, —NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH₃, —S(O)CH₃, —S(O)CH₂CH₃, —S(O)₂CH₃, —S(O)₂NH₂, —S(O)₂NHCH₃, —S(O)₂N(CH₃)₂, and —CH₂S(O)₂CH₃.
 2. The compound of claim 1 selected from Formulas Ia-n:


3. The compound of claim 1 wherein Formula Ia is selected from the structures:

where the A heterocyclyl ring is optionally substituted with one or more R⁵ groups.
 4. The compound of claim 1 wherein Formula If is selected from the structures:

where the A heterocyclyl ring is optionally substituted with one or more R⁵ groups.
 5. The compound of claim 1 wherein R⁴ is phenyl substituted with one or more groups selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —CN, —CF₃, —CH₂OH, —CO₂—, —CONH₂, —CONH(CH₃), —CON(CH₃)₂, —NO₂, —NH₂, —NHCH₃, —NHCOCH₃, —OH, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —SH, —NHC(═O)NHCH₃, —NHC(═O)NHCH₂CH₃, —NHS(O)₂CH₃, —N(CH₃)C(═O)OC(CH₃)₃, and —S(O)₂CH₃.
 6. The compound of claim 1 wherein R⁴ is an optionally substituted bicyclic heteroaryl group selected from 1H-indazole, 1H-indole, indolin-2-one, 1-(indolin-1-yl)ethanone, 1H-benzo[d][1,2,3]triazole, 1H-pyrazolo[3,4-b]pyridine, 1H-pyrazolo[3,4-d]pyrimidine, 1H-benzo[d]imidazole, 1H-benzo[d]imidazol-2(3H)-one, 1H-pyrazolo[3,4-e]pyridine, 1H-pyrrolo[2,3-e]pyridine, 3H-imidazo[4,5-e]pyridine, 7H-pyrrolo[2,3-d]pyrimidine, 7H-purine, 1H-pyrazolo[4,3-d]pyrimidine, 5H-pyrrolo[3,2-d]pyrimidine, 2-amino-1H-purin-6(9H)-one, quinoline, quinazoline, quinoxaline, isoquinoline, isoquinolin-1(2H)-one, 3,4-dihydroisoquinolin-(2H)-one, 3,4-dihydroquinolin-2(1H)-one, quinazolin-2(1H)-one, quinoxalin-2(1H)-one, 1,8-naphthyridine, pyrido[3,4-d]pyrimidine, and pyrido[3,2-b]pyrazine.
 7. The compound of claim 1 wherein optionally substituted R⁴ is selected from:

where the wavy line indicates the site of attachment.
 8. The compound of claim 7 wherein R⁴ is 1H-indazol-4-yl.
 9. The compound of claim 1 wherein R⁴ is an optionally substituted monocyclic heteroaryl group selected from 2-furanyl, 3-furanyl, 2-imidazolyl, 4-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-pyrazolyl, 4-pyrazolyl, 2-pyrazinyl, 3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl, 2-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrrolyl, 3-pyrrolyl, 2-thienyl, 3-thienyl, 5-tetrazolyl, 1-tetrazolyl, 2-tetrazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 3-triazolyl, and 1-triazolyl.
 10. The compound of claim 9 wherein R⁴ is 2-aminopyrimidin-5-yl.
 11. The compound of claim 1 wherein optionally substituted R⁴ is selected from:

where the wavy line indicates the site of attachment.
 12. The compound of claim 1 wherein one or more R⁵ groups are independently selected from F, Cl, Br, I, —CH₃, —CH₂CH₃, —C(CH₃)₃, —CH₂OH, —CH₂CH₂OH, —C(CH₃)₂OH, —CH₂OCH₃, —CN, —CH₂F, —CHF₂, —CF₃, —CO₂H, —COCH₃, —COC(CH₃)₃, —CO₂CH₃, —CONH₂, —CONHCH₃, —CON(CH₃)₂, —C(CH₃)₂CONH₂, —NO₂, —NH₂, —NHCH₃, —N(CH₃)₂, —NHCOCH₃, —NHS(O)₂CH₃, —N(CH₃)C(CH₃)₂CONH₂, —N(CH₃)CH₂CH₂S(O)₂CH₃, ═O, —OH, —OCH₃, —S(O)₂N(CH₃)₂, —SCH₃, —S(O)₂CH₃, cyclopropyl, cyclobutyl, oxetanyl, morpholino, and 1,1-dioxo-thiopyran-4-yl.
 13. The compound of claim 1 wherein two geminal R⁵ groups form cyclopropyl, cyclobutyl, cyclopentyl, tetrahydrofuryl, tetrahydropyranyl, oxetanyl, azetidinyl, pyrrolidinyl, piperidyl, piperazinyl, cyclohexyl, morpholino, or 1,1-dioxo-thiopyran-4-yl.
 14. The compound of claim 1 wherein mor is


15. A compound of claim 1 selected from: 1-[4-(3a,8-dimethyl-7-morpholin-4-yl-3,3a,8,8a-tetrahydro-2h-1-oxa-4,6,8-triaza-cyclopenta[a]inden-5-yl)-phenyl]-3-ethyl-urea; 5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)-4-methylpyrimidin-2-amine; 5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; 5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)-4-(trifluoromethyl)pyridyl-2-amine; 5-(4-morpholino-8,9-dihydro-7h-[1,3]oxazino[2,3-e]purin-2-yl)pyrimidin-2-amine; 5-(4-morpholino-6,7,8,9-tetrahydropyrido[2,1-e]purin-2-yl)pyrimidin-2-amine; 5-(4-morpholino-6,7,8,9-tetrahydropyrido[2,1-e]purin-2-yl)pyridin-2-amine; 5-(4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)-4-(trifluoromethyl)pyridyl-2-amine; 5-(4-morpholino-7,8-dihydro-6h-pyrrolo[2,1-e]purin-2-yl)pyrimidin-2-amine; 6,6-dimethyl-4-morpholino-2-(1H-pyrrolo[2,3-b]pyridin-5-yl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyridin-2-amine; 5-(4-morpholino-8,9-dihydrospiro[[1,3]oxazino[2,3-e]purine-7,1′-cyclopropane]-2-yl)pyrimidin-2-amine; 5-(4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; 5-(4-morpholino-8,9-dihydrospiro[[1,4]oxazino[3,4-e]purine-6,3′-oxetane]-2-yl)pyrimidin-2-amine; 5-(7,7-dimethyl-4-morpholino-8,9-dihydro-7h-[1,3]oxazino[2,3-e]purin-2-yl)pyrimidin-2-amine; 5-(4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyridin-2-amine; 5-(6,6-(hexadeuterio)dimethyl-4-morpholino-8,9-dihydro-6h-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; (S)-5-(6-ethyl-6-methyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; 5-(6,6,9-trimethyl-4-morpholino-6h-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; (R)-5-(6-ethyl-6-methyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; 5-(1-morpholin-4-yl-5,6,8a,9-tetrahydro-8h-7,10-dioxa-2,4,4b-triaza-phenanthren-3-yl)-pyrimidin-2-yl amine; 5-((S)-6-Morpholin-4-yl-2,3,3a,4-tetrahydro-1H-5-oxa-7,9,9b-triaza-cyclopenta[a]naphthalen-8-yl)-pyrimidin-2-ylamine; 4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)aniline; 1-(4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)-3-methylurea; 6,6-dimethyl-4-morpholino-2-(1H-pyrazol-4-yl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyri din-2-amine; 6,6-dimethyl-2-(1-methyl-1H-pyrazol-4-yl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 3-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenol; 2-(1H-indazol-5-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 6,6-dimethyl-2-(2-(4-methylpiperazin-1-yl) yridine-4-yl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; N-(2-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)methanesulfonamide; 6,6-dimethyl-4-morpholino-2-(6-morpholinopyridin-3-yl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 2-(1-benzyl-1H-pyrazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 2-(2-isopropoxypyridin-3-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; N-(2-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)acetamide; 2-(3,5-dimethyl-1H-pyrazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyridine-2-ol; 6-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyridine-3-amine; (R)-5-(4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine (S)-5-(4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; 2-(1-ethyl-1H-pyrazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)-N,N-dimethylbenzamide; tert-butyl 4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl(methyl)carbamate; 2-(3-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)acetonitrile; 6,6-dimethyl-4-morpholino-2-(3-morpholinophenyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 6,6-dimethyl-4-morpholino-2-(3-(morpholinomethyl)phenyl)-8,9-dihydro-6-[1,4]oxazino[3,4-e]purine; 2-(3-(benzyloxy)phenyl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 2-(1-isobutyl-1H-pyrazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 6,6-dimethyl-2-(6-(4-methylpiperazin-1-yl) yridine-3-yl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 2-(1H-indazol-4-yl)-6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)benzonitrile; 5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)nicotinamide; 5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)-N-methylpicolinamide; 2-(4-(benzyloxy)phenyl)-6,6-dimeth yl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 3-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)-N,N-dimethylaniline; 6,6-dimethyl-2-(4-(4-methylpiperazin-1-yl)phenyl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; 6,6-dimethyl-4-morpholino-2-(4-(piperidin-1-yl)phenyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine; N-(5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyridine-2-yl)acetamide; 5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)picolinamide; 6-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyridine-3-ol; (4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)(4-methylpiperazin-1-yl)methanone; N-cyclopropyl-3-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)benzamide; 5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)-N,N-dimethylpyrazin-2-amine; 1-(4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)-3-ethylurea; 1-(4-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenyl)-3-isopropyl urea; (2-(2-aminopyrimidin-5-yl)-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purine-6,6-diyl)dimethanol; 2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholino-8,9-dihydropyrazino[2,1-e]purin-6(7H)-one; 5-(8,8-Dimethyl-1-morpholin-4-yl-5,8-dihydro-6H-7-oxa-9-thia-2,4-diaza-fluoren-3-yl)-pyrimidin-2-ylamine; 2-(1H-indazol-4-yl)-4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6-[1,4]oxazino[3,4-e]purine; 3-(4-morpholino-6-(trifluoromethyl)-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)phenol; 5-(4-((2S,6R)-2,6-dimethylmorpholino)-6,6-dimethyl-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; 5-(4-(2,2-dimethylmorpholino)-6,6-dimethyl-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; N-(5-(6,6-dimethyl-4-morpholino-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-yl)acetamide; 5-(4-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl)-6,6-dimethyl-8,9-dihydro-6H-[1,4]oxazino[3,4-e]purin-2-yl)pyrimidin-2-amine; 2-(2-aminopyrimidin-5-yl)-6-methyl-4-morpholino-6,7-dihydropyrazino[2,1-e]purin-8(9H)-one; 5-(6,7-dimethyl-4-morpholino-6,7,8,9-tetrahydropyrazino[2,1-e]purin-2-yl)pyrimidin-2-amine; and 5-(8,8-Dimethyl-1-morpholin-4-yl-5,6-dihydro-8H-7-oxa-2,4,4b-triaza-fluoren-3-yl)-pyrimidin-2-ylamine.
 16. A pharmaceutical composition comprised of a compound of claim 1 and a pharmaceutically acceptable carrier, glidant, diluent, or excipient.
 17. The pharmaceutical composition according to claim 16, further comprising an additional therapeutic agent selected from a chemotherapeutic agent, an anti-inflammatory agent, an immunomodulatory agent, a neurotropic factor, an agent for treating cardiovascular disease, an agent for treating liver disease, an anti-viral agent, an agent for treating blood disorders, an agent for treating diabetes, and an agent for treating immunodeficiency disorders.
 18. A method of treating cancer in a patient comprised of administering to said patient a therapeutically effective amount of a compound of claim 1 wherein the cancer is breast, ovary, cervix, prostate, testis, genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, non-small cell lung carcinoma (NSCLC), small cell carcinoma, lung adenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidney carcinoma, renal, pancreatic, myeloid disorders, lymphoma, hairy cells, buccal cavity, naso-pharyngeal, pharynx, lip, tongue, mouth, small intestine, colon-rectum, large intestine, rectum, brain and central nervous system, Hodgkin's or leukemia.
 19. The method of claim 18 wherein the cancer is a brain cancer.
 20. The method of claim 18 further comprising administering to the patient an additional therapeutic agent selected from a chemotherapeutic agent, an anti-angiogenesis therapeutic agent, an anti-inflammatory agent, an immunomodulatory agent, a neurotropic factor, an agent for treating cardiovascular disease, an agent for treating liver disease, an anti-viral agent, an agent for treating blood disorders, an agent for treating diabetes, and an agent for treating immunodeficiency disorders.
 21. The method of claim 20 wherein the additional therapeutic agent is bevacizumab.
 22. A process for making a pharmaceutical composition which comprises combining a compound of claim 1 with a pharmaceutically acceptable carrier.
 23. A kit for treating a PI3K-mediated condition, comprising: a) a first pharmaceutical composition comprising a compound of claim 1; and b) instructions for use. 